Updated on 2024/03/20

写真a

 
MUNEYUKI Eiro
 
Organization
Faculty of Science and Engineering Professor
Other responsible organization
Physics Course of Graduate School of Science and Engineering, Master's Program
Physics Course of Graduate School of Science and Engineering, Doctoral Program
Contact information
The inquiry by e-mail is 《here
External link

Degree

  • 理学博士 ( 東京大学 )

  • 理学修士 ( 東京大学 )

Education

  • 1989.3
     

    The University of Tokyo   Graduate School, Division of Science   doctor course   completed

  • 1989.3
     

    The University of Tokyo   Graduate School, Division of Science   doctor course   completed

  • 1986.3
     

    The University of Tokyo   Graduate School, Division of Science   master course   completed

  • 1984.3
     

    The University of Tokyo   Faculty of Science   graduated

  • 1980.3
     

    洛星ヴィアトール学園高等学校   graduated

Research History

  • 2009.4 - 2020.3

    Gakushuin University   理学部物理学科   非常勤講師

  • 2008.4 -  

    Chuo University   理工学部   教授

  • 2007.4 - 2008.3

    Chuo University   理工学部   准教授

  • 2005.4 - 2007.3

    Chuo University   理工学部   助教授

  • 2004.4 - 2005.3

    Waseda University   教育学部   非常勤講師

  • 1990.3 - 2005.3

    Tokyo Institute of Technology   資源化学研究所   助手

  • 1991.4 - 1993.3

    Chiba University   園芸学部   非常勤講師

  • 1989.4 - 1990.3

    Saitama University   工学部   助手

▼display all

Professional Memberships

  • 日本生化学会

  • 日本生物物理学会

  • 日本物理学会

Research Interests

  • Biophysics

  • Biological energy transduction

Research Areas

  • Life Science / Biophysics  / 生物物理学

Papers

  • Conformational change mechanisms driving the rotation of F1-ATPase Reviewed

    Masahiro Motohashi, Mao Oide, Chigusa Kobayashi, Jaewoon Jung, Eiro Muneyuki, Yuji Sugita

    BioNano 8   8   2023.6

     More details

    Language:English  

    researchmap

  • Tight Chemomechanical Coupling of the F1 Motor Relies on Structural Stability. Reviewed International journal

    Mana Tanaka, Tomohiro Kawakami, Tomoaki Okaniwa, Yohei Nakayama, Shoichi Toyabe, Hiroshi Ueno, Eiro Muneyuki

    Biophysical journal   119 ( 1 )   48 - 54   2020.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    The F1 motor is a rotating molecular motor that ensures a tight chemomechanical coupling between ATP hydrolysis/synthesis reactions and rotation steps. However, the mechanism underlying this tight coupling remains to be elucidated. In this study, we used electrorotation in single-molecule experiments using an F1βE190D mutant to demonstrate that the stall torque was significantly smaller than the wild-type F1, indicating a loose coupling of this mutant, despite showing similar stepping torque as the wild-type. Experiments on the ATPase activity after heat treatment and gel filtration of the α3β3-subcomplex revealed the unstable structure of the βE190D mutant. Our results suggest that the tight chemomechanical coupling of the F1 motor relies on the structural stability of F1. We also discuss the difference between the stepping torque and the stall torque.

    DOI: 10.1016/j.bpj.2020.04.039

    PubMed

    researchmap

  • Characterization of Conformational Ensembles of Protonated N-glycans in the Gas-Phase Reviewed

    Re S, Watabe S, Nishima W, Muneyuki E, Yamaguchi Y, MacKerell AD Jr, Sugita Y

    Scientific Reports   8   1644   2018.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Nature Publishing Group  

    Ion mobility mass spectrometry (IM-MS) is a technique capable of investigating structural changes of biomolecules based on their collision cross section (CCS). Recent advances in IM-MS allow us to separate carbohydrate isomers with subtle conformational differences, but the relationship between CCS and atomic structure remains elusive. Here, we characterize conformational ensembles of gas-phase N-glycans under the electrospray ionization condition using molecular dynamics simulations with enhanced sampling. We show that the separation of CCSs between isomers reflects folding features of N-glycans, which are determined both by chemical compositions and protonation states. Providing a physicochemical basis of CCS for N-glycans helps not only to interpret IM-MS measurements but also to estimate CCSs of complex glycans.

    DOI: 10.1038/s41598-018-20012-0

    Web of Science

    researchmap

  • Simultaneous Formation and Spatial Patterning of ZnO on ITO Surfaces by Local Laser-Induced Generation of Microbubbles in Aqueous Solutions of [Zn(NH3)4]2+ Reviewed

    Sho Fujii, Ryuta Fukano, Yoshihito Hayami, Hiroaki Ozawa, Eiro Muneyuki, Noboru Kitamura, Masa-aki Haga

    ACS Appl. Mater. Interfaces   9   8413 - 8419   2017.2

     More details

    Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:American Chemical Society  

    researchmap

  • Crystal structures of the ATP-binding and ADP-release dwells of the V1 rotary motor. Reviewed

    Suzuki K, Mizutani K, Maruyama S, Shimono K, Imai FL, Muneyuki E, Kakinuma Y, Ishizuka-Katsura Y, Shirouzu M, Yokoyama S, Yamato I, Murata T

    Nat Commun.   27 ( 7 )   13235   2016.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:nature  

    researchmap

  • Formation of M-Like Intermediates in Proteorhodopsin in Alkali Solutions (pH >= similar to 8.5) Where the Proton Release Occurs First in Contrast to the Sequence at Lower pH Reviewed

    Jun Tamogami, Keitaro Sato, Sukuna Kurokawa, Takumi Yamada, Toshifumi Nara, Makoto Demura, Seiji Miyauchi, Takashi Kikukawa, Eiro Muneyuki, Naoki Kamo

    BIOCHEMISTRY   55 ( 7 )   1036 - 1048   2016.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER CHEMICAL SOC  

    Proteorhodopsin (PR) is an outward light-driven proton pump observed in marine eubacteria. Despite many structural and functional similarities to bacteriorhodopsin (BR) in archaea, which also acts as an outward proton pump, the mechanism of the photoinduced proton release and uptake is different between two H+-pumps. In this study, we investigated the pH dependence of the photocycle and proton transfer in PR reconstituted with the phospholipid membrane under alkaline conditions. Under these conditions, as the medium pH increased, a blue-shifted photoproduct (defined as M-a), which is different from M, with a pK(a) of ca. 9.2 was produced. The sequence of the photoinduced proton uptake and release during the photocycle was inverted with the increase in pH. A pK(a) value of ca. 9.5 was estimated for this inversion and was in good agreement with the pK(a) value of the formation of M-a (similar to 9.2). In addition, we measured the photoelectric current generated by PRs attached to a thin polymer film at varying pH. Interestingly, increases in the medium pH evoked bidirectional photocurrents, which may imply a possible reversal of the direction of the proton movement at alkaline pH. On the basis of these findings, a putative photocycle and proton transfer scheme in PR under alkaline pH conditions was proposed.

    DOI: 10.1021/acs.biochem.5b01196

    Web of Science

    researchmap

  • Hidden Entropy Production in F1-ATPase

    Nakayama Yohei, Tanaka Mana, Muneyuki Eiro

    Meeting Abstracts of the Physical Society of Japan   71   2645 - 2645   2016

     More details

    Language:Japanese   Publisher:The Physical Society of Japan  

    DOI: 10.11316/jpsgaiyo.71.2.0_2645

    researchmap

  • Single molecule thermodynamics of ATP synthesis by F-1-ATPase Reviewed

    Shoichi Toyabe, Eiro Muneyuki

    NEW JOURNAL OF PHYSICS   17   15008   2015.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:IOP PUBLISHING LTD  

    F0F1-ATP synthase is a factory for synthesizing ATP in virtually all cells. Its core machinery is the subcomplex F-1-motor (F-1-ATPase) and performs the reversible mechanochemical coupling. The isolated F-1-motor hydrolyzes ATP, which is accompanied by unidirectional rotation of its central gamma-shaft. When a strong opposing torque is imposed, the gamma-shaft rotates in the opposite direction and drives the F-1-motor to synthesize ATP. This mechanical-to-chemical free-energy transduction is the final and central step of the multistep cellular ATP-synthetic pathway. Here, we determined the amount of mechanical work exploited by the F-1-motor to synthesize an ATP molecule during forced rotations using a methodology combining a nonequilibrium theory and single molecule measurements of responses to external torque. We found that the internal dissipation of the motor is negligible even during rotations far from a quasistatic process.

    DOI: 10.1088/1367-2630/17/1/015008

    Web of Science

    researchmap

  • Torque Generation ofEnterococcus hiraeV-ATPase Reviewed

    Hiroshi Ueno, Yoshihiro Minagawa, Mayu Hara, Suhaila Rahman, Ichiro Yamato, Eiro Muneyuki, Hiroyuki Noji, Takeshi Murata, Ryota Iino

    Journal of Biological Chemistry   289 ( 45 )   31212 - 31223   2014.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:American Society for Biochemistry & Molecular Biology (ASBMB)  

    DOI: 10.1074/jbc.m114.598177

    researchmap

  • CHARMM Force-Fields with Modified Polyphosphate Parameters Allow Stable Simulation of the ATP-Bound Structure of Ca2+-ATPase Reviewed

    Yasuaki Komuro, Suyong Re, Chigusa Kobayashi, Eiro Muneyuki, Yuji Sugita

    JOURNAL OF CHEMICAL THEORY AND COMPUTATION   10 ( 9 )   4133 - 4142   2014.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER CHEMICAL SOC  

    Adenosine triphosphate (ATP) is an indispensable energy source in cells. In a wide variety of biological phenomena like glycolysis, muscle contraction/relaxation, and active ion transport, chemical energy released from ATP hydrolysis is converted to mechanical forces to bring about large-scale conformational changes in proteins. Investigation of structurefunction relationships in these proteins by molecular dynamics (MD) simulations requires modeling of ATP in solution and ATP bound to proteins with accurate force-field parameters. In this study, we derived new force-field parameters for the triphosphate moiety of ATP based on the high-precision quantum calculations of methyl triphosphate. We tested our new parameters on membrane-embedded sarcoplasmic reticulum Ca2+-ATPase and four soluble proteins. The ATP-bound structure of Ca2+-ATPase remains stable during MD simulations, contrary to the outcome in shorter simulations using original parameters. Similar results were obtained with the four ATP-bound soluble proteins. The new force-field parameters were also tested by investigating the range of conformations sampled during replica-exchange MD simulations of ATP in explicit water. Modified parameters allowed a much wider range of conformational sampling compared with the bias toward extended forms with original parameters. A diverse range of structures agrees with the broad distribution of ATP conformations in proteins deposited in the Protein Data Bank. These simulations suggest that the modified parameters will be useful in studies of ATP in solution and of the many ATP-utilizing proteins.

    DOI: 10.1021/ct5004143

    Web of Science

    researchmap

  • 3P154 Rotor-Stator Interactions in V1 and Vo from Enterococcus hirae V-ATPase(Molecular motor,Poster,The 52th Annual Meeting of the Biophysical Society of Japan(BSJ2014))

    Hiroshi Ueno, Yoshihiro Minagawa, Mayu Hara, Ichiro Yamato, Hiroyuki Noji, Takeshi Murata, Ryota Iino, Eiro Muneyuki

    Seibutsu Butsuri   54 ( 1 )   S274   2014

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.54.S274_4

    researchmap

  • 1P105 Molecular dynamics simulations of ATP/ADP bound forms of SR Ca^<2+>-ATPase using CHARMM force field with modified polyphosphate parameters(03. Membrane proteins,Poster,The 52nd Annual Meeting of the Biophysical Society of Japan(BSJ2014))

    Komuro Yasuaki, Re Suyong, Kobayashi Chigusa, Muneyuki Eiro, Sugita Yuji

    Seibutsu Butsuri   54 ( 1 )   S158   2014

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.54.S158_3

    researchmap

  • Thermodynamic Analyses of Nucleotide Binding to an Isolated Monomeric beta Subunit and the alpha(3)beta(3)gamma Subcomplex of F-1-ATPase Reviewed

    Yohsuke Kikuchi, Yusuke Naka, Hidemitsu Osakabe, Tetsuaki Okamoto, Tomoko Masaike, Hiroshi Ueno, Shoichi Toyabe, Eiro Muneyuki

    BIOPHYSICAL JOURNAL   105 ( 11 )   2541 - 2548   2013.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:CELL PRESS  

    Rotation of the gamma subunit of the F-1-ATPase plays an essential role in energy transduction by F-1-ATPase. Hydrolysis of an ATP molecule induces a 120 degrees step rotation that consists of an 80 degrees substep and 40 degrees substep. ATP binding together with ADP release causes the first 80 degrees step rotation. Thus, nucleotide binding is very important for rotation and energy transduction by F-1-ATPase. In this study, we introduced a beta Y341W mutation as an optical probe for nucleotide binding to catalytic sites, and a beta E190Q mutation that suppresses the hydrolysis of nucleoside triphosphate (NTP). Using a mutant monomeric beta Y341W subunit and a mutant alpha(3)beta(3)gamma subcomplex containing the beta Y341W mutation with or without an additional bE190Q mutation, we examined the binding of various NTPs (i.e., ATP, GTP, and ITP) and nucleoside diphosphates (NDPs, i.e., ADP, GDP, and IDP). The affinity (1/K-d) of the nucleotides for the isolated beta subunit and third catalytic site in the subcomplex was in the order ATP/ ADP &gt; GTP/ GDP &gt; ITP/IDP. We performed van't Hoff analyses to obtain the thermodynamic parameters of nucleotide binding. For the isolated b subunit, NDPs and NTPs with the same base moiety exhibited similar Delta H-0 and Delta G(0) values at 25 degrees C. The binding of nucleotides with different bases to the isolated b subunit resulted in different entropy changes. Interestingly, NDP binding to the alpha(3)beta(Y341W)(3)gamma subcomplex had similar K-d and Delta G(0) values as binding to the isolated beta(Y341W) subunit, but the contributions of the enthalpy term and the entropy term were very different. We discuss these results in terms of the change in the tightness of the subunit packing, which reduces the excluded volume between subunits and increases water entropy.

    DOI: 10.1016/j.bpj.2013.10.018

    Web of Science

    researchmap

  • Basic Properties of Rotary Dynamics of the Molecular Motor Enterococcus hirae V-1-ATPase Reviewed

    Yoshihiro Minagawa, Hiroshi Ueno, Mayu Hara, Yoshiko Ishizuka-Katsura, Noboru Ohsawa, Takaho Terada, Mikako Shirouzu, Shigeyuki Yokoyama, Ichiro Yamato, Eiro Muneyuki, Hiroyuki Noji, Takeshi Murata, Ryota Iino

    JOURNAL OF BIOLOGICAL CHEMISTRY   288 ( 45 )   32700 - 32707   2013.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Background: The chemomechanical coupling scheme of the rotary motor V-1-ATPase is incompletely understood. Results:Enterococcus hirae V-1-ATPase (EhV(1)) showed 120 degrees steps of rotation without substeps, as commonly seen with F-1-ATPase. Conclusion: The basic properties of rotary dynamics of EhV(1) are similar to those of Thermus thermophilus V-1-ATPase. Significance: This study revealed the common properties of V-1-ATPases as rotary molecular motors, distinct from those of F-1-ATPases.
    V-ATPases are rotary molecular motors that generally function as proton pumps. We recently solved the crystal structures of the V-1 moiety of Enterococcus hirae V-ATPase (EhV(1)) and proposed a model for its rotation mechanism. Here, we characterized the rotary dynamics of EhV(1) using single-molecule analysis employing a load-free probe. EhV(1) rotated in a counterclockwise direction, exhibiting two distinct rotational states, namely clear and unclear, suggesting unstable interactions between the rotor and stator. The clear state was analyzed in detail to obtain kinetic parameters. The rotation rates obeyed Michaelis-Menten kinetics with a maximal rotation rate (V-max) of 107 revolutions/s and a Michaelis constant (K-m) of 154 m at 26 degrees C. At all ATP concentrations tested, EhV(1) showed only three pauses separated by 120 degrees/turn, and no substeps were resolved, as was the case with Thermus thermophilus V-1-ATPase (TtV(1)). At 10 m ATP (?K-m), the distribution of the durations of the ATP-waiting pause fit well with a single-exponential decay function. The second-order binding rate constant for ATP was 2.3 x 10(6) m(-1) s(-1). At 40 mm ATP (?K-m), the distribution of the durations of the catalytic pause was reproduced by a consecutive reaction with two time constants of 2.6 and 0.5 ms. These kinetic parameters were similar to those of TtV(1). Our results identify the common properties of rotary catalysis of V-1-ATPases that are distinct from those of F-1-ATPases and will further our understanding of the general mechanisms of rotary molecular motors.

    DOI: 10.1074/jbc.M113.506329

    Web of Science

    researchmap

  • Properties of the electrogenic activity of bacteriorhodopsin. Reviewed International journal

    Shizuma Miyazaki, Makoto Matsumoto, Søren Bo Brier, Toshihiro Higaki, Takumi Yamada, Tetsuaki Okamoto, Hiroshi Ueno, Shoichi Toyabe, Eiro Muneyuki

    European biophysics journal : EBJ   42 ( 4 )   257 - 65   2013.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    In this study, we analyzed the photoelectric current generated by bacteriorhodopsin adsorbed on a polymer film, "Lumirror" (Muneyuki et al. in FEBS Lett 427:109-114, 1998). We could examine the photoelectric current over a wide range of light intensity and pH values using the same membrane owing to the mechanical and chemical stability of the thin polymer film. We analyzed the photoelectric current by comparison with a simple equivalent electric circuit. Analysis of experimental results obtained at different light intensities suggested that the electromotive force of the bacteriorhodopsin was independent of light intensity. The pH dependence of the photoelectric current suggested that the bacteriorhodopsin could generate a maximum electromotive force at approximately pH 6.

    DOI: 10.1007/s00249-012-0870-0

    PubMed

    researchmap

  • Free Energy Analysis on the Tom20-Presequence Complex in Solution for Understanding a Dynamic-Equilibrium Model Reviewed

    Komuro Yasuaki, Miyashita Naoyuki, Mori Takaharu, Muneyuki Eiro, Saitoh Takashi, Kohda Daisuke, Sugita Yuji

    BIOPHYSICAL JOURNAL   104 ( 2 )   665A   2013.1

  • 3P314 Conformational analysis of PA-glycans by replica-exchange molecular dynamics simulations(30.Miscellaneous topics,Poster,The 51st Annual Meeting of the Biophysical Society of Japan)

    Watanabe Shigehisa, Re Suyong, Muneyuki Eiro, Sugita Yuji

    Seibutsu Butsuri   53 ( 1 )   S264   2013

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.53.S264_1

    researchmap

  • 3P111 Molecular dynamics simulations of SR Ca^<2+>-ATPase using improved ATP force field(03. Membrane proteins,Poster)

    Komuro Yasuaki, Kobayashi Chigusa, Re Suyong, Muneyuki Eiro, Sugita Yuji

    Seibutsu Butsuri   53 ( 1 )   S230   2013

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.53.S230_3

    researchmap

  • Energetics of the presequence-binding poses in mitochondrial protein import through Tom20. Reviewed

    Komuro Y, Miyashita N, Mori T, Muneyuki E, Saitoh T, Kohda D, Sugita Y

    J Phys Chem B   117 ( 10 )   2864 - 2871   2013

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    researchmap

  • Recovery of state-specific potential of molecular motor from single-molecule trajectory Reviewed

    Shoichi Toyabe, Hiroshi Ueno, Eiro Muneyuki

    EPL   97 ( 4 )   40004   2012.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:EPL ASSOCIATION, EUROPEAN PHYSICAL SOCIETY  

    We have developed a novel method to evaluate the potential profile of a molecular motor at each chemical state from only the probe's trajectory and applied it to a rotary molecular motor F1-ATPase. By using this method, we could also obtain the information regarding the mechanochemical coupling and energetics. We demonstrate that the position-dependent transition of the chemical states is the key feature for the highly efficient free-energy transduction by F1-ATPase. Copyright (C) EPLA, 2012

    DOI: 10.1209/0295-5075/97/40004

    Web of Science

    researchmap

  • 1PS022 Effect of external torque on the rotation of TFβ E190D mutant(The 50th Annual Meeting of the Biophysical Society of Japan)

    Kawakami Tomohiro, Toyabe Shoici, Ueno Hiroshi, Kudo Seishi, Muneyuki Eiro

    Seibutsu Butsuri   52   S77 - S78   2012

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.52.S77_5

    researchmap

  • 2PT139 Molecular dynamics simulations of SR Ca2+-pump in the ATP bound forms(The 50th Annual Meeting of the Biophysical Society of Japan)

    Komuro Yasuaki, Kobayashi Chigusa, Muneyuki Eiro, Sugita Yuji

    Seibutsu Butsuri   52   S128   2012

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.52.S128_5

    researchmap

  • Thermodynamic efficiency and mechanochemical coupling of F-1-ATPase Reviewed

    Shoichi Toyabe, Takahiro Watanabe-Nakayama, Tetsuaki Okamoto, Seishi Kudo, Eiro Muneyuki

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   108 ( 44 )   17951 - 17956   2011.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:NATL ACAD SCIENCES  

    F-1-ATPase is a nanosized biological energy transducer working as part of FoF1-ATP synthase. Its rotary machinery transduces energy between chemical free energy and mechanical work and plays a central role in the cellular energy transduction by synthesizing most ATP in virtually all organisms. However, information about its energetics is limited compared to that of the reaction scheme. Actually, fundamental questions such as how efficiently F-1-ATPase transduces free energy remain unanswered. Here, we demonstrated reversible rotations of isolated F-1-ATPase in discrete 120 degrees steps by precisely controlling both the external torque and the chemical potential of ATP hydrolysis as a model system of FoF1-ATP synthase. We found that the maximum work performed by F-1-ATPase per 120 degrees step is nearly equal to the thermodynamical maximum work that can be extracted from a single ATP hydrolysis under a broad range of conditions. Our results suggested a 100% free-energy transduction efficiency and a tight mechanochemical coupling of F-1-ATPase.

    DOI: 10.1073/pnas.1106787108

    Web of Science

    researchmap

  • 理解したいということ

    宗行 英朗

    生物物理   51 ( 4 )   159 - 159   2011.7

     More details

    Language:Japanese   Publisher:一般社団法人 日本生物物理学会  

    DOI: 10.2142/biophys.51.159

    CiNii Books

    researchmap

  • Fabrication and Placement of a Ring Structure of Nanoparticles by a Laser-Induced Micronanobubble on a Gold Surface Reviewed

    Sho Fujii, Katsuhiko Kanaizuka, Shoichi Toyabe, Katsuaki Kobayashi, Eiro Muneyuki, Masa-aki Haga

    LANGMUIR   27 ( 14 )   8605 - 8610   2011.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER CHEMICAL SOC  

    We have developed a new fabrication method for a ring structure of assembled nanopartides on a gold surface by the use of continuous Nd:YAG laser light. A micronanobubble on a gold surface, created by laser local heating, acts as a template for the formation of the ring structure. Both Marangoni convection flow and capillary flow around the micronanobubble are responsible for the driving force to assemble nanoparticles such as CdSe Q-dots into the ring structure from the solution. Because a single micronanobubble was generated by the Nd:YAG laser focusing point, the precise positioning of the ring structure was feasible directly on the gold surface, which makes it possible to fabricate various patterns of rings such as arrays and letters and even a double-ring structure without any photomasks or any templates.

    DOI: 10.1021/la201616s

    Web of Science

    researchmap

  • Direct Observation of the Myosin Va Recovery Stroke That Contributes to Unidirectional Stepping along Actin Reviewed

    Katsuyuki Shiroguchi, Harvey F. Chin, Diane E. Hannemann, Eiro Muneyuki, Enrique M. De La Cruz, Kazuhiko Kinosita

    PLOS BIOLOGY   9 ( 4 )   e1001031   2011.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:PUBLIC LIBRARY SCIENCE  

    Myosins are ATP-driven linear molecular motors that work as cellular force generators, transporters, and force sensors. These functions are driven by large-scale nucleotide-dependent conformational changes, termed "strokes"; the "power stroke" is the force-generating swinging of the myosin light chain-binding "neck" domain relative to the motor domain "head" while bound to actin; the "recovery stroke" is the necessary initial motion that primes, or "cocks," myosin while detached from actin. Myosin Va is a processive dimer that steps unidirectionally along actin following a "hand over hand" mechanism in which the trailing head detaches and steps forward similar to 72 nm. Despite large rotational Brownian motion of the detached head about a free joint adjoining the two necks, unidirectional stepping is achieved, in part by the power stroke of the attached head that moves the joint forward. However, the power stroke alone cannot fully account for preferential forward site binding since the orientation and angle stability of the detached head, which is determined by the properties of the recovery stroke, dictate actin binding site accessibility. Here, we directly observe the recovery stroke dynamics and fluctuations of myosin Va using a novel, transient caged ATP-controlling system that maintains constant ATP levels through stepwise UV-pulse sequences of varying intensity. We immobilized the neck of monomeric myosin Va on a surface and observed real time motions of bead(s) attached site-specifically to the head. ATP induces a transient swing of the neck to the post-recovery stroke conformation, where it remains for similar to 40 s, until ATP hydrolysis products are released. Angle distributions indicate that the post-recovery stroke conformation is stabilized by &gt;= 5 k(B)T of energy. The high kinetic and energetic stability of the post-recovery stroke conformation favors preferential binding of the detached head to a forward site 72 nm away. Thus, the recovery stroke contributes to unidirectional stepping of myosin Va.

    DOI: 10.1371/journal.pbio.1001031

    Web of Science

    researchmap

  • 3G1446 Molecular dynamics simulation of outer mitochondrial membrane protein Tom20-presequence complex(3G Protein: Structure 4,The 49th Annual Meeting of the Biophysical Society of Japan)

    Komuro Yasuaki, Mori Takaharu, Muneyuki Eiro, Saitoh Takashi, Kohda Daisuke, Sugita Yuji

    Seibutsu Butsuri   51   S131 - S132   2011

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.51.S131_6

    researchmap

  • 2Q1636 The fast proton release of proteorhodopsin at low pH(Photobiology: Vision & Photoreception 2,The 48th Annual Meeting of the Biophysical Society of Japan)

    Tamogami Jun, Kikukawa Takashi, Shimono Kazumi, Nara Toshifumi, Muneyuki Eiro, Kamo Naoki

    Seibutsu Butsuri   51   S103   2011

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.51.S103_4

    researchmap

  • Experimental demonstration of information-to-energy conversion and validation of the generalized Jarzynski equality Reviewed

    Shoichi Toyabe, Takahiro Sagawa, Masahito Ueda, Eiro Muneyuki, Masaki Sano

    NATURE PHYSICS   6 ( 12 )   988 - 992   2010.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:NATURE PUBLISHING GROUP  

    In 1929, Leo Szilard invented a feedback protocol(1) in which a hypothetical intelligence-dubbed Maxwell's demon-pumps heat from an isothermal environment and transforms it into work. After a long-lasting and intense controversy it was finally clarified that the demon's role does not contradict the second law of thermodynamics, implying that we can, in principle, convert information to free energy(2-6). An experimental demonstration of this information-to-energy conversion, however, has been elusive. Here we demonstrate that a non-equilibrium feedback manipulation of a Brownian particle on the basis of information about its location achieves a Szilard-type information-to-energy conversion. Using real-time feedback control, the particle is made to climb up a spiral-staircase-like potential exerted by an electric field and gains free energy larger than the amount of work done on it. This enables us to verify the generalized Jarzynski equality(7), and suggests a new fundamental principle of an 'information-to-heat engine' that converts information into energy by feedback control.

    DOI: 10.1038/NPHYS1821

    Web of Science

    researchmap

  • Observation of DNA pinning at laser focal point on Au surface and its application to single DNA nanowire and cross-wire formation Reviewed

    Sho Fujii, Katsuaki Kobayashi, Katsuhiko Kanaizuka, Tetsuaki Okamoto, Shoichi Toyabe, Eiro Muneyuki, Masa-aki Haga

    BIOELECTROCHEMISTRY   80 ( 1 )   26 - 30   2010.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE SA  

    We report a new technique for fabricating a single DNA nanowire at a desired position in a sequential manner using the micronanobubble generated by laser local heating at the Au/water interface. In our previous report, we found the reversible pull-in/shrinkage of one end immobilized DNA strands near a Nd: YAG laser focal point on an Au surface. In further experiments, the pinning of DNA strands in the stretched state was observed on the Au surface only when the bubble has touched the free end of DNA. This pinning phenomenon was observed even on the alkane thiol modified Au surface as self-assembled monolayers (SAMs) such as hexanethiol, mercaptohexanol, and hexadecanethiol. However, no pinning was observed on the bovine serum albumin (BSA) modified surface. Since optical tweezers can manipulate a DNA modified bead (radius = 1.87 mu m), the bead was firstly fixed on a solid surface by being compressed with the optical tweezers, and the pulling and pinning of DNA on the bead were achieved. As a consequence, the laser local heating on the Au surface enables us to control the number and position of the one end immobilized DNA strands as DNA nanowires. (C) 2010 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.bioelechem.2010.04.004

    Web of Science

    researchmap

  • Nonequilibrium energetics of a single F1-ATPase molecule. Reviewed

    Toyabe S, Okamoto T, Watanabe-Nakayama T, Taketani H, Kudo S, Muneyuki E

    Physical Review Letters   104 ( 19 )   198103   2010.5

     More details

    Publisher:American Physical Society  

    DOI: 10.1103/PhysRevLett.104.198103

    PubMed

    researchmap

  • Chemo-Mechanical Coupling in F-1-ATPase Revealed by Catalytic Site Occupancy during Catalysis Reviewed

    Rieko Shimo-Kon, Eiro Muneyuki, Hiroshi Sakai, Kengo Adachi, Masasuke Yoshida, Kazuhiko Kinosita

    BIOPHYSICAL JOURNAL   98 ( 7 )   1227 - 1236   2010.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:CELL PRESS  

    F-1-ATPase is a rotary molecular motor in which the central gamma subunit rotates inside a cylinder made of alpha(3)beta(3) subunits. To clarify how ATP hydrolysis in three catalytic sites cooperate to drive rotation, we measured the site occupancy, the number of catalytic sites occupied by a nucleotide, while assessing the hydrolysis activity under identical conditions. The results show hitherto unsettled timings of ADP and phosphate releases: starting with ATP binding to a catalytic site at an ATP-waiting gamma angle defined as 0 degrees, phosphate is released at similar to 200 degrees, and ADP is released during quick rotation between 240 degrees and 320 degrees that is initiated by binding of a third ATP. The site occupancy remains two except for a brief moment after the ATP binding, but the third vacant site can bind a medium nucleotide weakly.

    DOI: 10.1016/j.bpj.2009.11.050

    Web of Science

    researchmap

  • Manipulation of Single DNA Using a Micronano Bubble Formed by Local Laser Heating on an Au-coated Surface Reviewed

    Sho Fujii, Katsuaki Kobayashi, Katsuhiko Kanaizuka, Tetsuaki Okamoto, Shoichi Toyabe, Eiro Muneyuki, Masa-aki Haga

    Chemistry Letters   39 ( 2 )   92 - 93   2010.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:The Chemical Society of Japan  

    We report a method for manipulating sequential single DNA strands by using a micronanobubble formed by local laser heating on a Au surface. DNA strands near the laser focal point were quickly pulled toward the focal point. This DNA pull-in phenomenon can be explained by Marangoni convection due to a micronanobubble generated by laser heating on the Au surface. The thickness of Au film plays a crucial role for absorbing the IR laser light.

    DOI: 10.1246/cl.2010.92

    CiNii Books

    researchmap

  • Allosteric model of an ion pump Reviewed

    Eiro Muneyuki, Ken Sekimoto

    PHYSICAL REVIEW E   81 ( 1 )   011137   2010.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER PHYSICAL SOC  

    We present a simple model of a free-energy transducer made of allosterically coupled two ratchet subsystems. Each of the subsystems transports particles from one particle reservoir to another. The coupling of the subsystems imposes correlated transitions of the potential profiles of the two subsystems. As a result, a downhill flux in one subsystem with higher chemical-potential difference drives an uphill flux in the other subsystem with lower chemical-potential difference. The direction of the driven flux inverts depending on the direction of the driving flux. The ratio between the fluxes conveyed by the two subsystems is variable and nonstoichiometric. By selecting appropriate parameters, the maximum ratio of the driven flux to driving flux and maximum free-energy transducing efficiency reaches some 90 and 40%, respectively. At a stalled state, the driven flux vanishes while the driving flux remains finite. The allosteric model enables explicit analysis of the timing between binding-unbinding of particles and transitions of potential profile. The behavior of the model is similar to but different from that of the alternate access model, which is a biochemical model for active transport proteins. Our model works also as a regulatory system. We suggest that the correlated transitions of the subsystems (subunits or domains) through allosteric interaction are the origin of the diverse functions of the protein machineries.

    DOI: 10.1103/PhysRevE.81.011137

    Web of Science

    researchmap

  • 2P175 Application of Simple Dark-Field Microscopy with High spatiotemporal resolution(The 48th Annual Meeting of the Biophysical Society of Japan)

    Ueno Hiroshi, Iino Ryota, Tabata Kazuhito V., Sakakihara Shouichi, Eiro Muneyuki, Noji Hiroyuki

    Seibutsu Butsuri   50 ( 2 )   S113   2010

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.50.S113_2

    researchmap

  • 2P123 Molecular dynamics simulation of outer mitochondrial membrane protein Tom20-presequence complex(The 48th Annual Meeting of the Biophysical Society of Japan)

    Komuro Yasuaki, Mori Takaharu, Muneyuki Eiro, Saitoh Takashi, Kohda Daisuke, Sugita Yuji

    Seibutsu Butsuri   50 ( 2 )   S103 - S104   2010

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.50.S103_6

    researchmap

  • 1P191 1B1450 Angle distribution analysis of hybrid F1-ATPase indicates another sub-substep rotation between two catalytic dwells(Molecular motor,Oral Presentations,The 48th Annual Meeting of the Biophysical Society of Japan)

    Ariga Takayuki, Muneyuki Eiro, Yoshida Masasuke

    Seibutsu Butsuri   50 ( 2 )   S53   2010

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.50.S53_1

    researchmap

  • Modulation of nucleotide binding to the catalytic sites of thermophilic F-1-ATPase by the epsilon subunit: Implication for the role of the epsilon subunit in ATP synthesis Reviewed

    Taichi Yasuno, Eiro Muneyuki, Masasuke Yoshida, Yasuyuki Kato-Yamada

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   390 ( 2 )   230 - 234   2009.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Effect of epsilon subunit on the nucleotide binding to the catalytic sites of F-1-ATPase from the thermophilic Bacillus PS3 (TF1) has been tested by using alpha(3)beta(3)gamma and alpha(3)beta(3)gamma epsilon complexes of TF1 containing beta Tyr341 to Trp substitution. The nucleotide binding was assessed with fluorescence quenching of the introduced Trp. The presence of the E subunit weakened ADP binding to each catalytic site, especially to the highest affinity site. This effect was also observed when GDP or IDP was used. The ratio of the affinity of the lowest to the highest nucleotide binding sites had changed two orders of magnitude by the c subunit. The differences may relate to the energy required for the binding change in the ATP synthesis reaction and contribute to the efficient ATP synthesis. (C) 2009 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.bbrc.2009.09.092

    Web of Science

    researchmap

  • A tin oxide transparent electrode provides the means for rapid time-resolved pH measurements: application to photoinduced proton transfer of bacteriorhodopsin and proteorhodopsin. Reviewed

    Tamogami J, Kikukawa T, Miyauchi S, Muneyuki E, Kamo N

    Photochemistry and photobiology   85 ( 2 )   578 - 589   2009.3

  • Watching 'ankle' action of myosin V

    Shiroguchi, Katsuyuki, Chin, Harvey, Muneyuki, Eiro, De La Cruz, Enrique M, Kinosita, Kazuhiko, Jr

    BIOPHYSICAL JOURNAL   96 ( 3 )   546A - 546A   2009.2

     More details

    Language:English  

    researchmap

  • 1TA4-10 Single molecule kinetic analysis of the temperature sensitive dwell of F1-ATPase(The 47th Annual Meeting of the Biophysical Society of Japan)

    Okamoto Tetsuaki, Toyabe Shouichi, Muneyuki Eiro

    Seibutsu Butsuri   49   S30 - S31   2009

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.49.S30_6

    researchmap

  • 3P-127 Temperature dependence of nucleotide binding to β-subunit of F1-ATPase revealed by fluorescence measurement.(Molecular motor,The 47th Annual Meeting of the Biophysical Society of Japan)

    Naka Yusuke, Muneyuki Eiro, Osakabe Hidemitsu, Masaike Tomoko

    Seibutsu Butsuri   49   S172   2009

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.49.S172_4

    researchmap

  • 2TA4-03 Stall torque of F1-ATPase(The 47th Annual Meeting of the Biophysical Society of Japan)

    Toyabe Shoichi, Watanabe-Nakayama Takahiro, Okamoto Tetsuaki, Kudo Seishi, Muneyuki Eiro

    Seibutsu Butsuri   49   S43   2009

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.49.S43_4

    researchmap

  • 3P-126 Stepping rotation of F1-ATPase at 2 mM ATP in the presence of DMSO (dimethylsulfoxide).(Molecular motor,The 47th Annual Meeting of the Biophysical Society of Japan)

    Nishizuka Kazuhiro, Toyabe Shoiti, Okamoto Tetsuaki, Muneyuki Eiro

    Seibutsu Butsuri   49   S172   2009

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.49.S172_3

    researchmap

  • Temperature dependence of the rotation and hydrolysis activities of F-1-ATPase Reviewed

    Shou Furuike, Kengo Adachi, Naoyoshi Sakaki, Rieko Shimo-Kon, Hiroyasu Itoh, Eiro Muneyuki, Masasuke Yoshida, Kazuhiko Kinosita

    BIOPHYSICAL JOURNAL   95 ( 2 )   761 - 770   2008.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:BIOPHYSICAL SOC  

    F-1-ATPase, a water-soluble portion of the enzyme ATP synthase, is a rotary molecular motor driven by ATP hydrolysis. To learn how the kinetics of rotation are regulated, we have investigated the rotational characteristics of a thermophilic F-1-ATPase over the temperature range 4-50 degrees C by attaching a polystyrene bead (or bead duplex) to the rotor subunit and observing its rotation under a microscope. The apparent rate of ATP binding estimated at low ATP concentrations increased from 1.2 x 10(6) M-1 s(-1) at 4 degrees C to 4.3 x 10(7) M-1 s(-1) at 40 degrees C, whereas the torque estimated at 2 mM ATP remained around 40 pN.nm over 4-50 degrees C. The rotation was stepwise at 4 degrees C, even at the saturating ATP concentration of 2 mM, indicating the presence of a hitherto unresolved rate-limiting reaction that occurs at ATP-waiting angles. We also measured the ATP hydrolysis activity in bulk solution at 4-65 degrees C. F1-ATPase tends to be inactivated by binding ADP tightly. Both the inactivation and reactivation rates were found to rise sharply with temperature, and above 30 degrees C, equilibrium between the active and inactive forms was reached within 2 s , the majority being inactive. Rapid inactivation at high temperatures is consistent with the physiological role of this enzyme, ATP synthesis, in the thermophile.

    DOI: 10.1529/biophysj.107.123307

    Web of Science

    researchmap

  • Effect of external torque on the ATP-driven rotation of F-1-ATPase Reviewed

    Takahiro Watanabe-Nakayama, Shoichi Toyabe, Selshi Kudo, Shigeru Suglyama, Masasuke Yoshida, Eiro Muneyuki

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   366 ( 4 )   951 - 957   2008.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    F-1-ATPase is a rotary molecular motor powered by the torque generated by another rotary motor F-0 to synthesize ATP in vivo. Therefore elucidation of the behavior of F-1 under external torque is very important. Here, we applied controlled external torque by electrorotation and investigated the ATP-driven rotation for the first time. The rotation was accelerated by assisting torque and decelerated by hindering torque, but F-1 rarely showed rotations in the ATP synthesis direction. This is consistent with the prediction by models based on the assumption that the rotation is tightly coupled to ATP hydrolysis and synthesis. At low ATP concentrations (2 and 5 mu M), 120 degrees stepwise rotation was observed. Due to the temperature rise during experiment, quantitative interpretation of the data is difficult, but we found that the apparent rate constant of ATP binding clearly decreased by hindering torque and increased by assisting torque. (c) 2007 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.bbrc.2007.12.049

    Web of Science

    researchmap

  • 2P-180 The role of the axle in F_1-ATPase revealed by rotation of a mutant containing a twisted γ subunit(The 46th Annual Meeting of the Biophysical Society of Japan)

    Hirakawa Takayuki, Masaike Tomoko, Iai Kentaro, Muneyuki Eiro, Yoshida Masasuke, Nishizaka Takayuki

    Seibutsu Butsuri   48   S103   2008

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.48.S103_1

    researchmap

  • 3P-214 Electrogenic activity of bacteriorhodopsin adsorbed on Lumirror membrane(The 46th Annual Meeting of the Biophysical Society of Japan)

    Muneyuki Eiro, Higaki Toshihiro, Matsumoto Makoto, Yamada Takumi, Okamoto Tetsuaki, Toyabe Shoichi

    Seibutsu Butsuri   48   S160   2008

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.48.S160_5

    researchmap

  • 3P-162 How F1 catches ATP from the environment at low temperature(The 46th Annual Meeting of the Biophysical Society of Japan)

    Okamoto Tetsuaki, Toyabe Shouichi, Muneyuki Eiro

    Seibutsu Butsuri   48   S152   2008

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.48.S152_4

    researchmap

  • 1P-167 Three steps of GT mutant (the mutant of F_1) at high ATP concentrations(The 46th Annual Meeting of the Biophysical Society of Japan)

    Taketani Hiroshi, Watanabe-Nakayama Takahiro, Okamoto Tetsuaki, Toyabe Shoichi, Muneyuki Eiro

    Seibutsu Butsuri   48   S47   2008

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.48.S47_3

    researchmap

  • 1P-154 Watching 'ankle' action of myosin V in the absence of actin(The 46th Annual Meeting of the Biophysical Society of Japan)

    Shiroguchi Katsuyuki, Chin Harvey, Muneyuki Eiro, De La Cruz Enrique M., Kinosita Jr.

    Seibutsu Butsuri   48   S45   2008

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.48.S45_2

    researchmap

  • 2P-310 Measurement of Violation of Fluctuation Dissipation Theorem of Single Molecule F_1-ATPase(The 46th Annual Meeting of the Biophysical Society of Japan)

    Toyabe Shoichi, Watanabe-Nakayama Takahiro, Okamoto Tetsuaki, Taketani Hiroshi, Kudo Seishi, Sugiyama Shigeru, Muneyuki Eiro

    Seibutsu Butsuri   48   S122   2008

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.48.S122_7

    researchmap

  • 1P-163 Asymmetric rotation of the hybrid F_1 containing one mutant a subunit(The 46th Annual Meeting of the Biophysical Society of Japan)

    Ariga Takayuki, Muneyuki Eiro, Yoshida Masasuke

    Seibutsu Butsuri   48   S46   2008

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.48.S46_5

    researchmap

  • 3P-156 Temperature dependency of the nucleotide binding to F1-ATPase(The 46th Annual Meeting of the Biophysical Society of Japan)

    Osakabe Hidemitsu, Toyabe Shoichi, Okamoto Tetuaki, Muneyuki Eiro

    Seibutsu Butsuri   48   S151   2008

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.48.S151_5

    researchmap

  • F-1-ATPase rotates by an asymmetric, sequential mechanism using all three catalytic subunits Reviewed

    Takayuki Ariga, Eiro Muneyuki, Masasuke Yoshida

    NATURE STRUCTURAL & MOLECULAR BIOLOGY   14 ( 9 )   841 - 846   2007.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:NATURE PUBLISHING GROUP  

    F-1- ATPase, the catalytic part of FoF1- ATP synthase, rotates the central c subunit within the alpha(3)beta(3) cylinder in 120 degrees steps, each step consuming a single ATP molecule. However, how the catalytic activity of each beta subunit is coordinated with the other two beta subunits to drive rotation remains unknown. Here we show that hybrid F-1 containing one or two mutant beta subunits with altered catalytic kinetics rotates in an asymmetric stepwise fashion. Analysis of the rotations reveals that for any given beta subunit, the subunit binds ATP at 0 degrees, cleaves ATP at similar to 200 degrees and carries out a third catalytic event at similar to 320 degrees. This demonstrates the concerted nature of the F-1 complex activity, where all three beta subunits participate to drive each 120 degrees rotation of the c subunit with a 120 degrees phase difference, a process we describe as a 'sequential three- site mechanism'.

    DOI: 10.1038/nsmb1296

    Web of Science

    researchmap

  • カーボンナノチューブ表面で自己組織的に生ずるポリベプチドαヘリックス構造のβシート構造転移--AFMおよびTEMによる解析

    杉山 幸宏, 井上 雄嗣, 宗行 英朗, 羽田 肇, 藤本 正之

    静岡大学大学院電子科学研究科研究報告   28 ( 28 )   53 - 57   2007.3

     More details

    Language:Japanese   Publisher:静岡大学  

    CiNii Books

    researchmap

  • Single molecule energetics of F-1-ATPase motor Reviewed

    Eiro Muneyuki, Takahiro Watanabe-Nakayama, Tetsuya Suzuki, Masasuke Yoshida, Takayuki Nishizaka, Hiroyuki Noji

    BIOPHYSICAL JOURNAL   92 ( 5 )   1806 - 1812   2007.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:BIOPHYSICAL SOCIETY  

    Motor proteins are essential in life processes because they convert the free energy of ATP hydrolysis to mechanical work. However, the fundamental question on how they work when different amounts of free energy are released after ATP hydrolysis remains unanswered. To answer this question, it is essential to clarify how the stepping motion of a motor protein reflects the concentrations of ATP, ADP, and P-i in its individual actions at a single molecule level. The F, portion of ATP synthase, also called F-1-ATPase, is a rotary molecular motor in which the central gamma-subunit rotates against the alpha(3)beta(3) cylinder. The motor exhibits clear step motion at low ATP concentrations. The rotary action of this motor is processive and generates a high torque. These features are ideal for exploring the relationship between free energy input and mechanical work output, but there is a serious problem in that this motor is severely inhibited by ADP. In this study, we overcame this problem of ADP inhibition by introducing several mutations while retaining high enzymatic activity. Using a probe of attached beads, stepping rotation against viscous load was examined at a wide range of free energy values by changing the ADP concentration. The results showed that the apparent work of each individual step motion was not affected by the free energy of ATP hydrolysis, but the frequency of each individual step motion depended on the free energy. This is the first study that examined the stepping motion of a molecular motor at a single molecule level with simultaneous systematic control of Delta G(ATP). The results imply that microscopically defined work at a single molecule level cannot be directly compared with macroscopically defined free energy input.

    DOI: 10.1529/biophysj.106.097170

    Web of Science

    researchmap

  • 3P241 Photo-induced proton transfer of proteorhodopsin (PR) measured by a SnO_2 transparent electrode(Photobiology- vision and photoreception,Poster Presentations)

    Tamogami Jun, Kikukawa Takashi, Miyauchi Seiji, Muneyuki Eiro, Kamo Naoki

    Seibutsu Butsuri   47   S263   2007

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.47.S263_2

    researchmap

  • 3P157 Catalytic Site-Occupancy during ATP Hydrolysis of F_1-ATPase That Lacks Non-Catalytic Nucleotide Binding Site(Molecular motors,Poster Presentations)

    Shimo-Kon Rieko, Muneyuki Eiro, Adachi Kengo, Furuike Shou, Sakai Hiroshi, Yoshida Masasuke, Kinoshita Kazuhiko

    Seibutsu Butsuri   47   S242   2007

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.47.S242_2

    researchmap

  • 3P156 F_1-ATPase rotates by an asymmetric, sequential three-site mechanism(Molecular motors,Oral Presentations)

    Ariga Takayuki, Muneyuki Eiro, Yoshida Masasuke

    Seibutsu Butsuri   47   S242   2007

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.47.S242_1

    researchmap

  • AFM and TEM observations of alpha-helix to beta-sheet conformational change occurring on carbon nanotubes Reviewed

    Yukihiro Sugiyama, Yuji Inoue, Eiro Muneyuki, Hajime Haneda, Masayuki Fujimoto

    JOURNAL OF ELECTRON MICROSCOPY   55 ( 3 )   143 - 149   2006.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:OXFORD UNIV PRESS  

    Bacteriorhodopsin (BR), which is rich in alpha-helical structure, was spread onto water with single-wall carbon nanotubes (SCNTs). After a Langmuir trough was used to apply compressive surface pressure to maintain the alpha-helices monolayer of denatured BR, the composite films comprising alpha-helices and SCNTs were transferred horizontally onto substrates. Atomic force microscopy (AFM) and fluorescence microscopy observation suggested that alpha-helices in contact with SCNTs changed into beta-sheets. High-resolution transmission electron microscopy (HR-TEM) showed 0.54 nm periodicity characteristic of the turn of alpha-helical structure in the SCNTs-free alpha-helix monolayer region and showed the 0.70 nm periodicity of beta-sheet pleated structure in the region where SCNTs were covered with unfolded BR. Unique features of carbon nanotubes that trigger conformational changes of a protein were revealed.

    DOI: 10.1093/jmicro/dfl024

    Web of Science

    researchmap

  • Kinetic mechanism of quinol oxidation by cytochrome bd studied with ubiquinone-2 analogs Reviewed

    Y Matsumoto, E Muneyuki, D Fujita, K Sakamoto, H Miyoshi, M Yoshida, T Mogi

    JOURNAL OF BIOCHEMISTRY   139 ( 4 )   779 - 788   2006.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPANESE BIOCHEMICAL SOC  

    Cytochrome bd is a heterodimeric terminal ubiquinol oxidase of Escherichia coli under microaerophilic growth conditions. The oxidase activity shows sigmoidal concentration-dependence with low concentrations of ubiquinols, and a marked substrate inhibition with high concentrations of ubiquinol-2 analogs [Sakamoto, K., Miyoshi, H., Takegami, K., Mogi, T., Anraku, Y., and Iwamura H. (1996) J. Biol. Chem. 271, 29897-29902]. Kinetic analysis of the oxidation of the ubiquinol-2 analogs, where the 2- or 3-methoxy group has been substituted with an azido or ethoxy group, suggested that its peculiar enzyme kinetics can be explained by a modified ping-pong bi-bi mechanism with the formation of inactive binary complex FS in the one-electron reduced oxygenated state and inactive ternary complex (E2S)S-n on the oxidation of the second quinol molecule. Structure-function studies on the ubiquinol-2 analogs suggested that the 6-diprenyl group and the 3-methoxy group on the quinone ring are involved in the substrate inhibition. We also found that oxidized forms of ubiquinone-2 analogs served as weak noncompetitive inhibitors. These results indicate that the mechanism for the substrate oxidation by cytochrome bd is different from that of the heme-copper terminal quinol oxidase and is tightly coupled to dioxygen reduction chemistry.

    DOI: 10.1093/jb/mvj087

    Web of Science

    researchmap

  • An alternative reaction pathway of F-1-ATPase suggested by rotation without 80 degrees/40 degrees substeps of a sluggish mutant at low ATP Reviewed

    K Shimabukuro, E Muneyuki, M Yoshida

    BIOPHYSICAL JOURNAL   90 ( 3 )   1028 - 1032   2006.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:BIOPHYSICAL SOCIETY  

    F-1-ATPase, a water-soluble portion of F0F1-ATP synthase, is a rotary motor driven by ATP hydrolysis. The central gamma-subunit rotates in the alpha(3)beta(3) cylinder by repeating four stages of rotation: ATP-binding dwell, rapid 80 degrees substep rotation, catalytic dwell, and rapid 40 degrees substep rotation. In the catalytic dwell, at least two catalytic reactions occur-cleavage of the enzyme-bound ATP and presumably release of the hydrolyzed product(s) from the enzyme - but we found that a slow ATP cleavage mutant of F-1-ATPase from thermophilic Bacillus PS3 rotates at low ATP concentration without substeps and the catalytic dwell. Analysis indicates that in this alternative reaction pathway the two catalytic reactions occur during the preceding long ATP-binding dwell. Thus, F-1-ATPase can operate through (at least) two competing reaction pathways, not necessarily through a simple consecutive reaction.

    Web of Science

    researchmap

  • 2P193 Characterization of interaction between β- and γ-subunit in F_1-ATPase using twisted γ mutant(37. Molecular motor (II),Poster Session,Abstract,Meeting Program of EABS & BSJ 2006)

    Shibano Satoko, Muneyuki Eiro, Masaike Tomoko, Okada Kaoru, Yoshida Masasuke, Nishizaka Takayuki

    Seibutsu Butsuri   46 ( 2 )   S344   2006

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.46.S344_1

    researchmap

  • 1P284 The behavior of F_1 motor in the presense of the uniform external torque applied with electrorotation(9. Molecular motor (I),Poster Session,Abstract,Meeting Program of EABS & BSJ 2006)

    Nakayama-Watanabe Takahiro, Muneyuki Eiro, Kudo Seishi, Sugiyama Shigeru, Yoshida Masasuke

    Seibutsu Butsuri   46 ( 2 )   S217   2006

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.46.S217_4

    researchmap

  • Rotation of F1-ATPase Reviewed

    Eiro Muneyuki, Masasuke Yoshida

    Handbook of ATPases: Biochemistry, Cell Biology, Pathophysiology   261 - 287   2005.12

     More details

    Language:English   Publishing type:Part of collection (book)   Publisher:Wiley Blackwell  

    DOI: 10.1002/3527606122.ch10

    Scopus

    researchmap

  • One rotary mechanism for F1-ATPase over ATP concentrations from millimolar down to nanomolar Reviewed

    N Sakaki, R Shimo-Kon, K Adachi, H Itoh, S Furuike, E Muneyuki, M Yoshida, K Kinosita

    BIOPHYSICAL JOURNAL   88 ( 3 )   2047 - 2056   2005.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:BIOPHYSICAL SOCIETY  

    F-1-ATPase is a rotary molecular motor in which the central gamma-subunit rotates inside a cylinder made of alpha(3)beta(3)-subunits. The rotation is driven by ATP hydrolysis in three catalytic sites on the beta-subunits. How many of the three catalytic sites are filled with a nucleotide during the course of rotation is an important yet unsettled question. Here we inquire whether F-1 rotates at extremely low ATP concentrations where the site occupancy is expected to be low. We observed under an optical microscope rotation of individual F1 molecules that carried a bead duplex on the gamma-subunit. Time-averaged rotation rate was proportional to the ATP concentration down to 200 pM, giving an apparent rate constant for ATP binding of 2 x 10(7) M-1 s(-1). A similar rate constant characterized bulk ATP hydrolysis in solution, which obeyed a simple Michaelis-Menten scheme between 6 mM and 60 nM ATP. F-1 produced the same torque of similar to40 pN . nm at 2 mM, 60 nM, and 2 nM ATP. These results point to one rotary mechanism governing the entire range of nanomolar to millimolar ATP, although a switchover between two mechanisms cannot be dismissed. Below 1 nM ATP, we observed less regular rotations, indicative of the appearance of another reaction scheme.

    DOI: 10.1529/biophysj.104.054668

    Web of Science

    researchmap

  • The characteristics of the (alpha(VC)-C-371)(3)(beta(RC)-C-337)(3 gamma) double mutant subcomplex of the TF1-ATPase indicate that the catalytic site at the alpha(TP)-beta(TP) interface with bound MgADP in crystal structures of MF1 represents a catalytic site containing inhibitory MgADP Reviewed

    S Bandyopadhyay, E Muneyuki, WS Allison

    BIOCHEMISTRY   44 ( 7 )   2441 - 2448   2005.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER CHEMICAL SOC  

    In the MF1 crystal structure with the MgADP-fluoroaluminate complex bound to two catalytic sites [Menz, R. I., Walker, J. E., and Leslie, A. G. W. (2001) Cell 106, 331-341], the guanidinium of betaR(337) is within 2.9 Angstrom of the alpha-oxygen of alphaS(370) and 3.7 Angstrom of a methyl group of alphaV(371) at the alphaE-beta(HC) interface. To examine the functional role of this contact, the (alphaV(371)C)(3)(betaR(337)C)(3)gamma subcomplex of the TF1-ATPase was prepared and characterized. Steady state ATPase activity of the reduced double-mutant is 30% of that of the wild type. Inactivation of the double mutant containing empty catalytic sites or MgADP bound to one catalytic site with CuCl2 cross-linked two alpha-beta pairs, whereas a single alpha-beta pair cross-linked when at least two catalytic sites contained MgADP. The reduced double mutant hydrolyzed substoichiometric ATP 100-fold more rapidly than the enzyme containing two cross-linked alpha-beta pairs. Addition of AlCl3 and NaF to the reduced double mutant after incubation with stoichiometric MgADP or 200 muM MgADP irreversibly inactivated the steady state ATPase activity with rate constants of 1.5 x 10(-2) and 4.1 x 10(-2) min(-1), respectively. In contrast, addition of AlCl3 and NaF to the cross-linked enzyme after incubation with stoichiometric or 200 muM MgADP irreversibly inactivated ATPase activity with a common rate constant of similar to10(-4) min(-1). Correlation of these results with crystal structures of MF1 suggests that the catalytic site at the alpha(TP)-beta(TP) interface is loaded first upon addition of nucleotides to nucleoticle-depleted F-1-ATPases and that the catalytic site at the alpha(TP)-beta(TP) interface with bound MgADP in crystal structures represents a catalytic site containing inhibitory MgADP.

    DOI: 10.1021/bi047694z

    Web of Science

    researchmap

  • 2SA05Ratchet model, single molecule observation of F_1 motor, and energetics

    Muneyuki E.

    Seibutsu Butsuri   45   S18   2005

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.45.S18_2

    researchmap

  • 3P166 A mechanism for the ATPase activity of F_1-ATPase that lacks non catalytic sites

    Shimo-Kon R., Muneyuki E., Sakaki N., Adachi K., Fruike S., Sakai H., Yoshida M., Kinosita K. Jr.

    Seibutsu Butsuri   45   S245   2005

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.45.S245_2

    researchmap

  • Chemomechanical coupling in F-1-ATPase revealed by simultaneous observation of nucleotide kinetics and rotation Reviewed

    T Nishizaka, K Oiwa, H Noji, S Kimura, E Muneyuki, M Yoshida, K Kinosita

    NATURE STRUCTURAL & MOLECULAR BIOLOGY   11 ( 2 )   142 - 148   2004.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:NATURE PUBLISHING GROUP  

    F-1-ATPase is a rotary molecular motor in which unidirectional rotation of the central subunit is powered by ATP hydrolysis in three catalytic sites arranged 120degrees apart around gamma. To study how hydrolysis reactions produce mechanical rotation, we observed rotation under an optical microscope to see which of the three sites bound and released a fluorescent ATP analog. Assuming that the analog mimics authentic ATP, the following scheme emerges: (i) in the ATP-waiting state, one site, dictated by the orientation of gamma, is empty, whereas the other two bind a nucleotide; (ii) ATP binding to the empty site drives an similar to80degrees rotation of gamma; (iii) this triggers a reaction(s), hydrolysis and/or phosphate release, but not ADP release in the site that bound ATP one step earlier; (iv) completion of this reaction induces further 40degrees rotation.

    DOI: 10.1038/nsmb721

    Web of Science

    researchmap

  • Rotation of F-1-ATPase at very low ATP concentrations

    N Sakaki, R Shimo-Kon, H Itoh, K Adachi, S Furuike, E Muneyuki, M Yoshida, K Kinosita

    BIOPHYSICAL JOURNAL   86 ( 1 )   476A - 476A   2004.1

     More details

    Language:English   Publisher:BIOPHYSICAL SOCIETY  

    Web of Science

    researchmap

  • 3P147 Site occupancy and ATP hydrolysis activity of F_1-ATPase without noncatalytic nucleotide binding site

    Shimo-Kon R., Muneyuki E., Sakaki N., Adachi K., Furuike S., Itoh H., Yoshida M., Kinosita Jr. K.

    Seibutsu Butsuri   44   S226   2004

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.44.S226_3

    researchmap

  • 3P148 Single molecule analysis of the rotation in F_1-ATPase mutant: the effect of ATP-binding dwell length on the catatytic dwell

    Shimabukuro K., Muneyuki E., Yoshida M.

    Seibutsu Butsuri   44   S226   2004

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.44.S226_4

    researchmap

  • 3P145 Rotation and torque of F_1-ATPase at extremely low ATP concentrations

    Sakaki N., Shimo-Kon R., Itoh H., Adachi K., Muneyuki E., Yoshida M., Kinosita Jr. K., Furuike S.

    Seibutsu Butsuri   44   S226   2004

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.44.S226_1

    researchmap

  • Catalysis and rotation of F-1 motor: Cleavage of ATP at the catalytic site occurs in 1 ms before 40 degrees substep rotation Reviewed

    K Shimabukuro, R Yasuda, E Muneyuki, KY Hara, K Kinosita, M Yoshida

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   100 ( 25 )   14731 - 14736   2003.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:NATL ACAD SCIENCES  

    F-1, a water-soluble portion of FoF1-ATP synthase, is an ATIP hydrolysis-driven rotary motor. The central gamma-subunit rotates in the alpha(3)beta(3) cylinder by repeating the following four stages of rotation: ATP-binding dwell, rapid 801 substep rotation, interim dwell, and rapid 40degrees substep rotation. At least two 1-ms catalytic events occur in the interim dwell, but it is still unclear which steps in the ATPase cycle, except for ATIP binding, correspond to these events. To discover which steps, we analyzed rotations of F-1 subcomplex (alpha(3)beta(3)gamma) from thermophilic Bacillus PS3 under conditions where cleavage of ATIP at the catalytic site is decelerated: hydrolysis of ATP by the catalytic-site mutant F, and hydrolysis of a slowly hydrolyzable substrate ATPgammaS (adenosine 5'-[gamma-thio]triphosphate) by wild-type F-1. In both cases, interim dwells were extended as expected from bulk phase kinetics, confirming that cleavage of ATP takes place during the interim dwell. Furthermore, the results of ATPgammaS hydrolysis by the mutant F-1 ensure that cleavage of ATIP most likely corresponds to one of the two 1-ms events and not some other faster undetected event. Thus, cleavage of ATP on F-1 occurs in 1 ms during the interim dwell, and we call this interim dwell catalytic dwell.

    DOI: 10.1073/pnas.2434983100

    Web of Science

    researchmap

  • Origin of apparent negative cooperativity of F-1-ATPase Reviewed

    S Ono, KY Hara, J Hirao, B Matsui, H Noji, M Yoshida, E Muneyuki

    BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS   1607 ( 1 )   35 - 44   2003.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    In order to get insight into the origin of apparent negative cooperativity observed for F-1-ATPase, we compared ATPase activity and ATPMg binding of mutant subcomplexes of thermophilic F-1-ATPase, alpha((W463F))(3)beta((Y341W))(3)gamma and alpha((K175A/T176A/W463F)3)beta((Y341W)3)gamma. For alpha((W463F)3)beta((Y341W)3)gamma, apparent Km's of ATPase kinetics (4.0 and 233 muM) did not agree with apparent K-m's deduced from fluorescence quenching of the introduced tryptophan residue (on the order of nM, 0.016 and 13 muM). On the other hand, in case of alpha((K175A/T176A/W463F))(3)beta((Y341W)3)gamma, which lacks noncatalytic nucleotide binding sites, the apparent K., of ATPase activity (10 muM) roughly agreed with the highest K. of fluorescence measurements (27 muM). The results indicate that in case of alpha((W463F)3)beta((Y341W))(3)gamma, the activating effect of ATP binding to noncatalytic sites dominates overall ATPase kinetics and the highest apparent K,, of ATPase activity does not represent the ATP binding to a catalytic site. In case of alpha((K175A/T176A/W463F)3)beta((Y341W))(3)gamma, the K-m of ATPase activity reflects the ATP binding to a catalytic site due to the lack of noncatalytic sites. The Eadie-Hofstee plot of ATPase reaction by alpha((K175A/T176A/W463F)3)beta((Y341W)3)gamma was rather linear compared with that of alpha((W463F)3)beta((Y341W))(3)gamma, if not perfectly straight, indicating that the apparent negative cooperativity observed for wild-type F-1-ATPase is due to the ATP binding to catalytic sites and noncatalytic sites. Thus, the frequently observed K-m's of 100-300 muM and 1-30 muM range for wild-type F-1-ATPase correspond to ATP binding to a noncatalytic site and catalytic site, respectively. (C) 2003 Elsevier B.V. All rights reserved.

    DOI: 10.1016/j.bbabio.2003.08.006

    Web of Science

    researchmap

  • Evidence for rotation of V-1-ATPase Reviewed

    H Imamura, M Nakano, H Noji, E Muneyuki, S Ohkuma, M Yoshida, K Yokoyama

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   100 ( 5 )   2312 - 2315   2003.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:NATL ACAD SCIENCES  

    VoV1-ATPase is responsible for acidification of eukaryotic intracellular compartments and ATP synthesis of Archaea and some eubacteria. From the similarity to FoF1-ATP synthase, VoV1-ATPase has been assumed to be a rotary motor, but to date there are no experimental data to support this. Here we visualized the rotation of single molecules of V-1-ATPase, a catalytic subcomplex of VoV1-ATPase. V-1-ATPase from Thermus thermophilus was immobilized onto a glass surface, and a bead was attached to the D or IF subunit through the biotin-streptavidin linkage. In both cases we observed ATP-dependent rotations of beads, the direction of which was always counterclockwise viewed from the membrane side. Given that three ATP molecules are hydrolyzed per one revolution, rates of rotation agree consistently with rates of ATP hydrolysis at saturating ATP concentrations. This study provides experimental evidence that VoV1-ATPase is a rotary motor and that both D and F subunits constitute a rotor shaft.

    DOI: 10.1073/pnas.0436796100

    Web of Science

    researchmap

  • Measurement of site occupancy and rotation speed of F1-ATPase without noncatalytic nucleotide binding site

    Kon-Shimo R., Muneyuki E., Iai K., Sakaki N., Adachi K., Itoh H., Furuike S., Maki Y., Yoshida M., Kinosita Jr. K.

    Seibutsu Butsuri   43   S134   2003

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.43.S134_2

    researchmap

  • Relationship between the free energy of ATP hydrolysis and step rotation of F_1-ATPase

    Muneyuki E., Watanabe T., Noji H., Nishizaka T., Yoshida M.

    Seibutsu Butsuri   43   S134   2003

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.43.S134_3

    researchmap

  • Analysis of rotation mechanism of F_1-ATPase using twisted γ mutant

    Iai K., Muneyuki E., Yoshida M.

    Seibutsu Butsuri   43   S134   2003

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.43.S134_4

    researchmap

  • Single molecule analysis of cooperative rotation mechanism using hybrid F_1-ATPase

    Ariga T., Shimabukura K., Muneyuki E., Yoshida M.

    Seibutsu Butsuri   43   S133   2003

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.43.S133_3

    researchmap

  • Cl- concentration dependence of photovoltage generation by halorhodopsin from Halobacterium salinarum Reviewed

    E Muneyuki, C Shibazaki, Y Wada, M Yakushizin, H Ohtani

    BIOPHYSICAL JOURNAL   83 ( 4 )   1749 - 1759   2002.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:BIOPHYSICAL SOCIETY  

    The photovoltage generation by halorhodopsin from Halobacterium salinarum (shR) was examined by adsorbing shR-containing membranes onto a thin polymer film. The photovoltage consisted of two major components: one with a sub-millisecond range time constant and the other with a millisecond range time constant with different amplitudes, as previously reported. These components exhibited different Cl- concentration dependencies (0.1-9 M). We found that the time constant for the fast component was relatively independent of the Cl- concentration, whereas the time constant for the slow component increased sigmoidally at higher Cl- concentrations. The fast and the slow processes were attributed to charge (Cl-) movements within the protein and related to Cl- ejection, respectively. The laser photolysis studies of shR-membrane suspensions revealed that they corresponded to the formation and the decay of the N intermediate. The photovoltage amplitude of the slow component exhibited a distorted bell-shaped Cl- concentration dependence, and the Cl- concentration dependence of its time constant suggested a weak and highly cooperative Cl--binding site(s) on the cytoplasmic side (apparent K-D of similar to5 M and Hill coefficient greater than or equal to5). The Cl- concentration dependence of the photovoltage amplitude and the time constant for the slow process suggested a competition between spontaneous relaxation and ion translocation. The time constant for the relaxation was estimated to be &gt; 100 ms.

    Web of Science

    researchmap

  • F1-ATPase changes its conformations upon phosphate release. Reviewed International journal

    Masaike,T, Muneyuki,E, Noji,H, Kinosita,K.Jr, Yoshida,M

    J.Biol.Chem.   277 ( 24 )   21643 - 21649   2002.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:American Society for Biochemistry and Molecular Biology  

    Motor proteins, myosin, and kinesin have gamma-phosphate sensors in the switch II loop that play key roles in conformational changes that support motility. Here we report that a rotary motor, F1-ATPase, also changes its conformations upon phosphate release. The tryptophan mutation was introduced into Arg-333 in the beta subunit of F1-ATPase from thermophilic Bacillus PS3 as a probe of conformational changes. This residue interacts with the switch II loop (residues 308-315) of the beta subunit in a nucleotide-bound conformation. The addition of ATP to the mutant F1 subcomplex alpha3beta(R333W)3gamma caused transient increase and subsequent decay of the Trp fluorescence. The increase was caused by conformational changes on ATP binding. The rate of decay agreed well with that of phosphate release monitored by phosphate-binding protein assays. This is the first evidence that the beta subunit changes its conformation upon phosphate release, which may share a common mechanism of exerting motility with other motor proteins.

    PubMed

    researchmap

  • The presence of phosphate at a catalytic site suppresses the formation of the MgADP-inhibited form of F-1-ATPase Reviewed

    N Mitome, S Ono, T Suzuki, K Shimabukuro, E Muneyuki, M Yoshida

    EUROPEAN JOURNAL OF BIOCHEMISTRY   269 ( 1 )   53 - 60   2002.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:BLACKWELL PUBLISHING LTD  

    F-1-ATPase is inactivated by entrapment of MgADP in catalytic sites and reactivated by MgATP or P-i. Here, using a mutant alpha(3)beta(3)gamma complex of thermophilic F-1-ATPase (alphaW463F/betaY341W) and monitoring nucleotide binding by fluorescence quenching of an introduced tryptophan, we found that P-i interfered with the binding of MgATP to F-1-ATPase, but binding of MgADP was interfered with to a lesser extent. Hydrolysis of MgATP by F-1-ATPase during the experiments did not obscure the interpretation because another mutant, which was able to bind nucleotide but not hydrolyse ATP (alphaW463F/betaE190Q/betaY341W), also gave the same results. The half-maximal concentrations of P-i that suppressed the MgADP-inhibited form and interfered with MgATP binding were both approximate to 20 mM. It is likely that the presence of P-i at a catalytic site shifts the equilibrium from the MgADP-inhibited form to the enzyme-MgADP-P-i complex, an active intermediate in the catalytic cycle.

    Web of Science

    researchmap

  • 2O1515 Torque measurement of F1-motor using magnetic tweezers

    Noji H., Itoh H., Muneyuki E., Adachi K., Yoshida M., Kinoshita K.

    Seibutsu Butsuri   42 ( 2 )   S146   2002

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.42.S146_2

    researchmap

  • 2O1445 Rotation and site occupancy of F_1-ATPase at low ATP concentrations

    Sakaki N., Shimo R., Itoh H., Adachi K., Muneyuki E., Yoshida M., Kinosita K.

    Seibutsu Butsuri   42 ( 2 )   S145   2002

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.42.S145_4

    researchmap

  • 2O1400 A mutant F1-ATPase resistant to ADP inhibition (2)

    Muneyuki E., Suzuki T., Iai K., Noji H., Nishizaka T., Yoshida M

    Seibutsu Butsuri   42 ( 2 )   S145   2002

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.42.S145_1

    researchmap

  • 2O1500 Rotation analysis of F_1-ATPase mutant αзβ(E190D)зγ

    Shimabukuro K., Muneyuki E., Yoshida M.

    Seibutsu Butsuri   42 ( 2 )   S146   2002

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.42.S146_1

    researchmap

  • 2O1330 Origin of substeps in 120°-stepping rotation of F_1-ATPase

    Nishizaka T., Oiwa K., Kimura S., Kinosita K., Yoshida M., Muneyuki E., Noji H.

    Seibutsu Butsuri   42 ( 2 )   S144   2002

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.42.S144_3

    researchmap

  • ATP synthase - A marvellous rotary engine of the cell Reviewed

    M Yoshida, E Muneyuki, T Hisabori

    NATURE REVIEWS MOLECULAR CELL BIOLOGY   2 ( 9 )   669 - 677   2001.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:NATURE PUBLISHING GROUP  

    ATP synthase can be thought of as a complex of two motors - the ATP-driven Ft motor and the proton-driven F-o motor - that rotate in opposite directions. The mechanisms by which rotation and catalysis are coupled in the working enzyme are now being unravelled on a molecular scale.

    Web of Science

    researchmap

  • Alkane derivative-bacteriorhodopsin interaction: proton transport and protein structure Reviewed

    A Shibata, H Ikema, S Ueno, E Muneyuki, T Higuti

    COLLOIDS AND SURFACES B-BIOINTERFACES   22 ( 1 )   31 - 38   2001.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    The effects of alkane derivatives, 1-alcohols (ROH), aliphatic amine hydrochlorides (RNH2. HCl) and sodium aliphatic carboxylates (ROONa), on the proton pumping activity of bacteriorhodopsin (bR) in a purple membrane have been examined. Photocurrents in bR in the purple membrane adsorbed onto polyester thin film were recorded before and after exposure to these test substances. The peak photocurrent in bR was reversibly suppressed by each substance. From the dose-response curve, the concentrations required to reduce the peak capacitive current by 50% were determined for each homolog and then the standard free energies per CH2 for the adsorption of the alkane derivatives to the site of action were estimated: -3.13 kJ mol(-1) for ROH, -3.05 kJ mol(-1) for RNH3+, and -2.95 kJ mol(-1) for ROO-. The proton pumping activity of bR was mainly suppressed by the hydrophobic interaction with the additive. The relative potencies of the functional groups of the alkane derivatives were almost comparable between 1-octanol (C8OH) and octylamine hydrochloride (C8NH3+) and about 10 times less effective for sodium octanoate (C8OO-) than for others. The addition of C8OH or C8OO- changed the absorption spectra of bR with a maximum at 560 nm to the spectra of the intermediate state with a maximum at 480 nm, while the C8NH3+ decreased the intensity of the 560 nm band only with no blue-shift by the 480 nm band. We conclude that the action of the alkane derivatives is nonspecific and directed to all organized purple membrane structures and that the binding sites of the ROH and ROO- are different from that of RNH3+. (C) 2001 Elsevier Science B.V. All rights reserved.

    Web of Science

    researchmap

  • <視点/動く>分子が動く--新しい機能性材料アプローチ:熱ゆらぎの中で働くナノ分子モーター

    宗行 英朗, 吉田 賢右

    未来材料   1 ( 5 )   2 - 5   2001.5

     More details

    Language:Japanese   Publisher:エヌ・ティー・エス  

    CiNii Books

    researchmap

  • Pause and rotation of F1-ATPase during catalysis Reviewed

    Yoko Hirono-Hara, Hiroyuki Noji, Masaya Nishiura, Eiro Muneyuki, Kiyotaka Y.Hara, Ryohei Yasuda, Kazuhiko Kinosita,Jr, Masasuke Yoshida

    Proc.Natl. Acad.Sci.USA   98 ( 24 )   13649 - 13654   2001.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:National Academy of Science(US)  

    DOI: 10.1073/pnas.241365698

    researchmap

  • Rotation analysis of beta(E190D)F1-ATPase mutant.

    Shimabukuro K., Muneyuki E., Yasuda R., Hara K.Y., Kinosita K., Yoshida M.

    Seibutsu Butsuri   41   S199   2001

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.41.S199_1

    researchmap

  • Expression of the proton pump activity of bacteriorhodopsin with hydrophobic cations : the effect of tertiary amine derivatives

    Yorimitsu A., Shibata A., Ueno S., Higuchi T., Muneyuki E., Kamo N.

    Seibutsu Butsuri   41   S62   2001

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.41.S62_2

    researchmap

  • Analysis of ADP-Mg inhibition of F1-ATPase

    Hirono-Hara Y, Noji H, Muneyuki E, Kinosita K Jr, Yoshida M

    Seibutsu Butsuri   41   S198   2001

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.41.S198_3

    researchmap

  • Direct measurement of rotary potentials of F_1 motor using magnetic tweezers

    Noji H., Itoh H., Shio M., Adachi K., Muneyuki E., Yoshida M., Kinosita K.Jr..

    Seibutsu Butsuri   41   S199   2001

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.41.S199_4

    researchmap

  • Kinetic analysis about MgADP inhibition of F1-ATPase

    Kawashima K., Muneyuki E., Kinosita K.Jr

    Seibutsu Butsuri   41   S198   2001

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.41.S198_2

    researchmap

  • F0F1-ATP synthase: general structural features of 'ATP-engine' and a problem on free energy transduction Reviewed

    E Muneyuki, H Noji, T Amano, T Masaike, M Yoshida

    BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS   1458 ( 2-3 )   467 - 481   2000.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    Web of Science

    researchmap

  • Properties of the stochastic energization-relaxation channel model for vectorial ion transport Reviewed

    E Muneyuki, TA Fukami

    BIOPHYSICAL JOURNAL   78 ( 3 )   1166 - 1175   2000.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:BIOPHYSICAL SOCIETY  

    A model for the primary active transport by an ion pump protein is proposed. The model, the "energization-relaxation channel model," describes an ion pump as a multiion channel that undergoes stochastic transitions between two conformational states by external energy supply. When the potential profile along ion transport pathway is asymmetrical, a net ion flux is induced by the transitions. In this model, the coupling of the conformational change and ion transport is stochastic and loose. The model qualitatively reproduces known properties of active transport such as the effect of ion concentration gradient and membrane potential on the rate of transport and the inhibition of ion transport at high ion concentration. We further examined the effect of various parameters on the ion transport properties of this model. The efficiency of the coupling was almost 100% under some conditions.

    Web of Science

    researchmap

  • Rotation of F-1-ATPase and the hinge residues of the beta subunit Reviewed

    T Masaike, N Mitome, H Noji, E Muneyuki, R Yasuda, K Kinosita, M Yoshida

    JOURNAL OF EXPERIMENTAL BIOLOGY   203 ( 1 )   1 - 8   2000.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:COMPANY OF BIOLOGISTS LTD  

    Rotation of a motor protein, F-1-ATPase, was demonstrated using a unique single-molecule observation system. This paper reviews what has been clarified by this system and then focuses on the role of residues at the hinge region of the beta subunit, We have visualised rotation of a single molecule of F-1-ATPase by attaching a fluorescent actin filament to the top of the gamma subunit in the immobilised F-1-ATPase, thus settling a major controversy regarding the rotary catalysis, The rotation of the gamma subunit was exclusively in one direction, as could be predicted by the crystal structure of bovine heart F-1-ATPase, Rotation at low ATP concentrations revealed that one revolution consists of three 120 degrees steps, each fuelled by the binding of an ATP to the beta subunit, The mean work done by a 120 degrees step was approximately 80 pN nm, a value close to the free energy liberated by hydrolysis of one ATP molecule, implying nearly 100% efficiency of energy conversion. The torque is probably generated by the beta subunit, which undergoes large opening-closing domain motion upon binding of AT(D)P, We identified three hinge residues, beta His179, beta Gly180 and beta Gly181, whose peptide bond dihedral angles are drastically changed during domain motion. Simultaneous substitution of these residues with alanine resulted in nearly complete loss (99%) of ATPase activity. Single or double substitution of the two Gly residues did not abolish the ATPase activity. However, reflecting the shift of the equilibrium between the open and closed forms of the beta subunit, single substitution caused changes in the propensity to generate the kinetically trapped Mg-ADP inhibited form: Gly180Ala enhanced the propensity and Gly181Ala abolished the propensity. In spite of these changes, the mean rotational torque was not changed significantly for any of the mutants.

    Web of Science

    researchmap

  • A stochastic channel model for free energy transduction

    Muneyuki E., Fukami T.A.

    Seibutsu Butsuri   40   S208   2000

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.40.S208_3

    CiNii Books

    researchmap

  • alpha(3)beta(3)gamma complex of F-1-ATPase from thermophilic Bacillus PS3 can maintain steady-state ATP hydrolysis activity depending on the number of non-catalytic sites Reviewed

    T Amano, T Matsui, E Muneyuki, H Noji, K Hara, M Yoshida, T Hisabori

    BIOCHEMICAL JOURNAL   343 ( 1 )   135 - 138   1999.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:PORTLAND PRESS LTD  

    Homogeneous preparations of alpha(3)beta(3)gamma complexes with one, two or three non-competent non-catalytic site(s) were performed as described [Amano, Hisabori, Muneyuki, and Yoshida (1996) J. Biol. Chem. 271, 18128-18133] and their properties were compared with those of the wild-type complex. The ATPase activity of the complex with three non-competent non-catalytic sites decayed rapidly to an inactivated state, as reported previously [Matsui, Muneyuki, Honda, Allison, Dou, and Yoshida (1997) J. Biol. Chem. 272, 8215-8221]. In contrast, the complex with one or two non-competent non-catalytic sites displayed a substantial steady-state phase activity depending on the number of non-competent non-catalytic sites in the complex. This result indicates that one competent non-catalytic site can maintain the continuous catalytic turnover of the enzyme and can potentially relieve all three catalytic sites from inhibition by MgADP(-). Furthermore, the results suggest that the interaction between three non-catalytic sites might not be as strong as that between catalytic sites, which are all strictly required for a continuous catalytic turnover.

    Web of Science

    researchmap

  • The noncatalytic site-deficient alpha(3)beta(3)gamma subcomplex and F0F1-ATP synthase can continuously catalyse ATP hydrolysis when P-i is present Reviewed

    D Bald, E Muneyuki, T Amano, J Kruip, T Hisabori, M Yoshida

    EUROPEAN JOURNAL OF BIOCHEMISTRY   262 ( 2 )   563 - 568   1999.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:BLACKWELL SCIENCE LTD  

    We investigated ATP hydrolysis by a mutant (Delta NC) alpha(3)beta(3)gamma subcomplex of F0F1-ATP synthase from the thermophilic Bacillus PS3 that is defective in the noncatalytic nucleotide binding sites. This mutant subcomplex was activated by inorganic phosphate ions (P-i) and did not show continuous ATP hydrolysis activity in the absence of P-i. P-i also activated the wild-type alpha(3)beta(3)gamma subcomplex in a similar manner. Sulphate activated wild-type alpha(3)beta(3)gamma but not Delta NC alpha(3)beta(3)gamma, indicating that P-i activation did not involve noncatalytic sites but that sulphate activation did. P-i also activated ATP hydrolysis and coupled proton translocation by the wild-type and Delta NC F0F1-ATP synthases reconstituted into vesicle membranes.

    Web of Science

    researchmap

  • Chloride concentration dependency of the electrogenic activity of halorhodopsin Reviewed

    D Okuno, M Asaumi, E Muneyuki

    BIOCHEMISTRY   38 ( 17 )   5422 - 5429   1999.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER CHEMICAL SOC  

    The capacitive photoelectric current responses of the halorhodopsins from Halobacterium salinarum (shR) and from Natronobacterium pharaonis (phR) were studied using membrane fragments adsorbed onto a thin polyester film. The electric current of shR was not much affected by ionic strength or cations present in the medium (Na+, K+, Li+, Mg2+, or Ca2+), but was greatly influenced by the Cl- concentration. It increased biphasically as the Cl- concentration increased from 0 to 5 M, then decreased and almost vanished at around 10 or 12 M, Apparent K-d's of about 0.1 and 6 M were deduced for the Kd of Cl- uptake sites. We had to assume a sigmoidal increase of Cl- binding with a Will coefficient of about 8 at the cytoplasmic, Cl- release site(s), The half-maximum Cl- concentration for the sigmoidal binding was about 7.5 M. The electric current of phR had a maximum around 30 mM Cl- and biphasically decreased at higher Cl- concentrations. The apparent K-d for the Cl- uptake site was 5 mM. The biphasic decrease in the transport activity was explained by assuming a sum of simple hyperbolic type binding (K-d = 0.2 M) and sigmoidally increasing binding with a Hill coefficient of 10 on the cytoplasmic side. The half-maximum concentration of the latter cooperative binding was 5.6 M. This great difference between the apparent affinity of the release site of shR and that of phR can explain the previously reported difference between the Cl- dependency of their photocycles, These results also suggest that there may be multiple Cl- binding sites in the Cl- transport pathway. A simple sequence of Cl- transport steps based on a multiion channel model is proposed.

    Web of Science

    researchmap

  • Cross-linking of two beta subunits in the closed conformation in F-1-ATPase Reviewed

    SP Tsunoda, E Muneyuki, T Amano, M Yoshida, H Noji

    JOURNAL OF BIOLOGICAL CHEMISTRY   274 ( 9 )   5701 - 5706   1999.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    In the crystal structure of mitochondrial F-1-ATPase, two beta subunits with a bound Mg-nucleotide are in "closed" conformations, whereas the third beta subunit without bound nucleotide is in an "open" conformation. In this "CCO" (beta-closed beta-closed beta-open) conformational state, Ile-390s of the two closed beta subunits, even though they are separated by an intervening alpha subunit, have a direct contact. We replaced the equivalent Ile of the alpha(3)beta(3)gamma subcomplex of thermophilic F-1-ATPase with Cys and observed the formation of the beta-beta cross-link through a disulfide bond. The analysis of conditions required for the cross-link formation indicates that: (i) F-1-ATPase takes the CCO conformation when two catalytic sites are filled with Mg-nucleotide, (ii) intermediate(s) with the CCO conformation are generated during catalytic cycle, (iii) the Mg-ADP inhibited form is in the CCO conformation, and (iv) F-1-ATPase dwells in conformational state(s) other than CCO when only one (or none) of catalytic sites is filled by Mg-nucleotide or when catalytic sites are filled by Mg2+-free nucleotide, The alpha(3)beta(3)gamma subcomplex containing the beta-beta cross-link retained the activity of uni-site catalysis but lost that of multiple catalytic turnover, suggesting that open-closed transition of beta subunits is required for the rotation of gamma subunit but not for hydrolysis of a single ATP.

    Web of Science

    researchmap

  • Time-resolved measurements of photovoltage generation by bacteriorhodopsin and halorhodopsin adsorbed on a thin polymer film Reviewed

    E Muneyuki, C Shibazaki, H Ohtani, D Okuno, M Asaumi, T Mogi

    JOURNAL OF BIOCHEMISTRY   125 ( 2 )   270 - 276   1999.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPANESE BIOCHEMICAL SOC  

    We constructed a time-resolved photovoltage measurement system and examined the photovoltage kinetics of wild-type bacteriorhodopsin, its D96N mutant, and halorhodopsins from Halobacterium salinarum and Natronobacterium pharaonis. Upon illumination with a laser hash, wild-type bacteriorhodopsin showed photovoltage generation with fast (10-100 mu s range) and slow (ms range) components while D96N lacked the latter, as reported previously [Holz, M., Drachev, L.A., Mogi, T., Otto, H., Kaulen, A.D., Heyn, M.P., Skulachev, V.P., and Khorana, H.G. (1989) Proc. Natl. Acad. Sci. USA 86, 2167-2171]. In contrast, photovoltage generation in halorhodopsins from H. salinarum and N. pharaonis was significant only in the ms time range. On the basis of the photovoltage kinetics and photocycle, we conclude that major charge (chloride) movements within halorhodopsin occur during the formation and decay of the N intermediate in the ms range. These observations are discussed in terms of the "Energization-Relaxation Channel Model" [Muneyuki, E., Ikematsu, M., and Yoshida, M. (1996) J. Phys. Chem. 100, 19687-19691].

    Web of Science

    researchmap

  • Direct observation of the rotation of the mutant F1-ATPase without noncatalytic binding site

    Hirono Y., Nishiura M., Muneyuki E., Noji H., Kinoshita K., Yoshida M.

    Seibutsu Butsuri   39   S145   1999

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.39.S145_4

    CiNii Books

    researchmap

  • Crystallization of F1-ATPase α_3β_3γ sub-assembly

    Shirakihara Y., Kambara M., Amano T., Tsunoda S., Muneyuki E., Shirakihara C., Shiratori A., Yoshida M.

    Seibutsu Butsuri   39   S104   1999

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.39.S104_4

    CiNii Books

    researchmap

  • Excitation wavelength dependence of the fluorescence spectra of bacteriorhodopsin and halorhodopsin.

    Ohtani H., Muneyuki E.

    Seibutsu Butsuri   39   S73   1999

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.39.S73_1

    CiNii Books

    researchmap

  • V-ATPase from Thermus thermophilus is inactivated during ATP hydrolysis but can synthesis ATP. Reviewed

    Yokoyama, K, Muneyuki, E, Amano, T, Mizutani, S, Yoshida, M, Ohkuma, S

    The Journal of Biological Chemistry   273   12248 - 12252   1998.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1074/jbc.273.32.20504

    researchmap

  • V-ATPase of Thermus thermophilus is inactivated during ATP hydrolysis but can synthesize ATP Reviewed

    K Yokoyama, E Muneyuki, T Amano, S Mizutani, M Yoshida, M Ishida, S Ohkuma

    JOURNAL OF BIOLOGICAL CHEMISTRY   273 ( 32 )   20504 - 20510   1998.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    The ATP hydrolysis of the V-1-ATPase of Thermus thermophilus have been investigated with an ATP-regenerating system at 25 degrees C, The ratio of ATPase activity to ATP concentration ranged from 40 to 4000 mu M; from this, an apparent K-m of 240 +/- 24 mu M and a V-max of 5.2 +/-0.5 units/mg were deduced. An apparent negative cooperativity, which is frequently observed in case of F-1-ATPases, was not observed for the V-1-ATPase, Interestingly, the rate of hydrolysis decayed rapidly during ATP hydrolysis, and the ATP hydrolysis finally stopped. Furthermore, the inactivation of the V-1-ATPase was attained by a prior incubation with ADP-Mg. The inactivated V-1-ATPase contained 1.5 mol of ADP/mol of enzyme. Difference absorption spectra generated from addition of ATP-Mg to the isolated subunits revealed that the A subunit can bind ATP-Mg, whereas the B subunit cannot. The inability to bind ATP-Mg is consistent with the absence of Walker motifs in the B subunit, These results indicate that the inactivation of the V-1-ATPase during ATP hydrolysis is caused by entrapping inhibitory ADP-Mg in a catalytic site. Light-driven ATP synthesis by bacteriorhodopsin-VoV1-ATPase proteoliposomes was observed, and the rate of ATP synthesis was approximately constant. ATP synthesis occurred in the presence of an ADP-Mg of which concentration was high enough to induce complete inactivation of ATP hydrolysis of VoV1-ATPase, This result indicates that the ADP-Mg-inhibited form is not produced in ATP synthesis reaction.

    Web of Science

    researchmap

  • A new system for the measurement of electrogenicity produced by ion pumps using a thin polymer film: examination of wild type bacteriorhodopsin and the D96N mutant over a wide pH range Reviewed

    E Muneyuki, D Okuno, M Yoshida, A Ikai, H Arakawa

    FEBS LETTERS   427 ( 1 )   109 - 114   1998.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    We developed a new assay system for the measurement of capacitive electric currents generated by ion pumps using the thin polymer film 'Lumirror' (Toray Co., Japan), This system enables us to examine the electrogenicity of ion pumps over a wide range of experimental conditions with high reproducibility due to the mechanical and chemical stability, the high electric resistance and the high electric capacitance of the thin polymer film, Using this method, we examined the photoelectric response of wild type bacteriorhodopsin and its D96N mutant over a wide pH range (2.8-10.0), The results were explained in terms of the affinities of the proton binding sites for translocated protons. A possibility that the direction of the proton transfer from the Schiff base was influenced by the protonation/deprotonation state of the surrounding proton binding sites was suggested. We also found that this film can be used as a substrate for atomic force microscopy (AFM) samples and hence the active purple membrane was observed with AFM. (C) 1998 Federation of European Biochemical Societies.

    Web of Science

    researchmap

  • ATP synthesis by F0F1-ATP synthase independent of noncatalytic nucleotide binding sites and insensitive to azide inhibition Reviewed

    D Bald, T Amano, E Muneyuki, B Pitard, JL Rigaud, J Kruip, T Hisabori, M Yoshida, M Shibata

    JOURNAL OF BIOLOGICAL CHEMISTRY   273 ( 2 )   865 - 870   1998.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    ATP hydrolyzing activity of a mutant alpha(3) beta(3) gamma subcomplex of F0F1-ATP synthase (Delta NC) from the thermophilic Bacillus PS3, which lacked noncatalytic nucleotide binding sites, was inactivated completely soon after starting the reaction (Matsui, T., Muneyuki, E., Honda, M., Allison, W. S., Dou, C., and Yoshida, M. (1997) J. Biol. Chem. 272, 8215-8221), This inactivation is caused by rapid accumulation of the "MgADP inhibited form" which, in the case of wild-type enzyme, would be relieved by ATP binding to noncatalytic sites. We reconstituted F0F1-ATP synthase into liposomes together with bacteriorhodopsin and measured illumination-driven ATP synthesis. Remarkably, Delta NC F0F1-ATP synthase catalyzed continuous turnover of ATP synthesis while it could not promote ATP-driven proton translocation. ATP synthesis by Delta NC F0F1-ATP synthase, as well as wild-type enzyme, proceeded even in the presence of azide, an inhibitor of ATP hydrolysis that stabilizes the MgADP inhibited form. The time course of ATP synthesis by Delta NC F0F1-ATP synthase was linear, and gradual acceleration to the maximal rate, which was observed for the wild-type enzyme, was not seen. Thus, ATP synthesis can proceed without nucleotide binding to noncatalytic sites even though the rate is sub-maximal. These results indicate that the MgADP inhibited form is not produced in ATP synthesis reaction, and in this regard, ATP synthesis may not be a simple reversal of ATP hydrolysis.

    Web of Science

    researchmap

  • Thermophilic F-1-ATPase is activated without dissociation of an endogenous inhibitor, epsilon subunit Reviewed

    Y Kato, T Matsui, N Tanaka, E Muneyuki, T Hisabori, M Yoshida

    JOURNAL OF BIOLOGICAL CHEMISTRY   272 ( 40 )   24906 - 24912   1997.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Subunit complexes (alpha(3) beta(3) gamma, alpha(3) beta(3) gamma delta, alpha(3) beta(3) gamma epsilon, and alpha(3) beta(3) gamma delta epsilon) of thermophilic F-1-ATPase were prepared, and their catalytic properties were compared to know the role of delta and epsilon subunits in catalysis. The presence of delta subunit in the complexes had slight inhibitory effect on the ATPase activity, The effect of epsilon subunit was more profound, The (-epsilon) complexes, alpha(3) beta(3) gamma and alpha(3) beta(3) gamma delta, initiated ATP hydrolysis without a lag. In contrast, the (+epsilon) complexes, alpha(3) beta(3) gamma epsilon and alpha(3) beta(3) gamma delta epsilon, started hydrolysis of ATP (&lt;700 mu M) with a lag phase that was gradually activated during catalytic turnover. As ATP concentration increased, the lag phase of the (+epsilon) complexes became shorter, and it was not observed above 1 mM ATP, Analysis of binding and hydrolysis of the ATP analog, 2',3'-O-(2,4,6-trinitrophenyl) ATP, suggested that the (+epsilon) complexes bound substrate only slowly. Differing from Escherichia coli F-1-ATPase, the activation of the (+epsilon) complexes from the lag phase was not due to dissociation of epsilon subunit since the re-isolated activated complex retained epsilon subunit. This indicates that there are two alternative forms of the (+epsilon) complex, inhibited form and activated form, and the inhibited one is converted to the activated one during catalytic turnover.

    Web of Science

    researchmap

  • A single mutation at the catalytic site of TF1-alpha(3)beta(3)gamma complex switches the kinetics of ATP hydrolysis from negative to positive cooperativity Reviewed

    E Muneyuki, M Odaka, M Yoshida

    FEBS LETTERS   413 ( 1 )   55 - 59   1997.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    Previously, we reported the substitution of Tyr(341) of the F-1-ATPase beta subunit from a thermophilic Bacillus strain PS3 with leucine, cysteine,:or alanine (M, Odaka ct al, J. Biochem., 115 (1994) 789-796). These mutations resulted in a great decrease in the affinity of the isolated beta subunit for ATP-Mg and an increase in the apparent K-m of the alpha(3) beta(3) gamma complex in ATP hydrolysis when examined above 0.1 mM ATP. Here, we examined the ATPase activity of the mutant complexes in a wide range of ATP concentration and found that the mutants exhibited apparent positive cooperativity in ATP hydrolysis. This is the first clear demonstration that a single mutation in the catalytic sites converts the kinetics from apparent negative cooperativity in the wild-type alpha(3) beta(3) gamma complex to apparent positive cooperativity, The conversion of apparent cooperativity could be explained in terms of a simple kinetic scheme based on the binding change model proposed by Boyer, (C) 1997 Federation of European Biochemical Societies.

    Web of Science

    researchmap

  • Catalytic activity of the alpha(3)beta(3)gamma complex of F-1-ATPase without noncatalytic nucleotide binding site Reviewed

    T Matsui, E Muneyuki, M Honda, WS Allison, C Dou, M Yoshida

    JOURNAL OF BIOLOGICAL CHEMISTRY   272 ( 13 )   8215 - 8221   1997.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    A mutant alpha(3) beta(3) gamma complex of F-1-ATPase from thermophilic Bacillus PS3 was generated in which noncatalytic nucleotide binding sites lost their ability to bind nucleotides. It hydrolyzed ATP at an initial rate with cooperative kinetics (K-m(1), 4 mu M; K-m(2), 135 mu M) similar to the wild-type complex. However, the initial rate decayed rapidly to an inactivated form. Since the inactivated mutant complex contained 1.5 mol of ADP/mol of complex, this inactivation seemed to be caused by entrapping inhibitory MgADP in a catalytic site. Indeed, the mutant complex was nearly completely inactivated by a 10 min prior incubation with equimolar MgADP. Analysis of the progress of inactivation after initiation of ATP hydrolysis as a function of ATP concentration indicated that the inactivation was optimal at ATP concentrations in the range of K-m(1). In the presence of ATP, the wildtype complex dissociated the inhibitory [H-3]ADP pre-loaded onto a catalytic site whereas the mutant complex did not. Lauryl dimethylamineoxide promoted release of preloaded inhibitory [H-3]ADP in an ATP-dependent manner and partly restored the activity of the inactivated mutant complex. Addition of ATP promoted single-site hydrolysis of 2',3'-O-(2,4,6-trinitrophenyl)-ATP preloaded at a single catalytic site of the mutant complex. These results indicate that intact noncatalytic sites are essential for continuous catalytic turnover of the F-1-ATPase but are not essential for catalytic cooperativity of F-1-ATPase observed at ATP concentrations below similar to 300 mu M.

    Web of Science

    researchmap

  • The heterogeneous interaction of substoichiometric TNP-ATP and F-1-ATPase from Escherichia coli Reviewed

    E Muneyuki, T Hisabori, T Sasayama, K Mochizuki, M Yoshida

    JOURNAL OF BIOCHEMISTRY   120 ( 5 )   940 - 945   1996.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPAN BIOCHEMICAL SOC  

    The interactions of substoichiometric TNP-ATP and F-1-ATPase from Escherichia coli (EF(1)) were examined and compared with those in the case of mitochondrial F-1-ATPase (MF(1)) and F-1-ATPase from thermophilic Bacillus PS3 (TF1). EF(1) hydrolyzed substoichiometric TNP-ATP faster than TF1 or MF(1), although some 20% of the TNP-ATP remained unhydrolyzed even in the presence of excess chase ATP. The affinity of the catalytic site of EF(1) for the product, TNP-ADP, was weaker than that of TF1 or MF(1), and the TNP-ADP was readily released upon addition of excess ATP. The amplitude of the difference absorption spectrum induced by binding of TNP-AT(D)P to EF(1) was smaller than that of MF(1) or TF1 under similar experimental conditions. When an excess amount of TNP-ATP was added to EF(1) and the change of the difference spectrum was measured, the shape of the difference spectrum of the ATP-replaceable fraction was very similar to that in the case of binding of TNP-ATP to the isolated beta subunit of TF1, indicating that the rapidly replaceable fraction of bound TNP-ATP was actually at the catalytic site and most of the non-replaceable portion was bound at noncatalytic sites. Weaker affinity of the catalytic site for TNP-ATP may account for the heterogeneous binding and hydrolysis under the conditions described in this paper.

    Web of Science

    researchmap

  • ΔμH+ Dependency of proton translocation by bacteriorhodopsin and a stochastic energization-relaxation channel model Reviewed

    Muneyuki,E, Ikematsu,M, Yoshida,M

    J.Phys.Chem.   100 ( 50 )   19687 - 19691   1996.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:アメリカ化学会  

    researchmap

  • Catalytic activities of alpha(3)beta(3)gamma complexes of F-1-ATPase with 1, 2, or 3 incompetent catalytic sites Reviewed

    T Amano, T Hisabori, E Muneyuki, M Yoshida

    JOURNAL OF BIOLOGICAL CHEMISTRY   271 ( 30 )   18128 - 18133   1996.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    In order to know how many functional catalytic sites are necessary for ATPase activity of F-1-ATPase from a thermophilic Bacillus PS3, a new method of isolating homogeneous preparations of the alpha(3) beta(3) gamma complex with 1, 2, or 3 incompetent catalytic sites was developed. Ten glutamic acids (Glu . Tag) were linked to the C terminus of the catalytically incompetent beta(E190Q) subunit. The Glu Tag itself did not affect ATPase activity of the complexes. Two kinds of alpha(3) beta(3) gamma complexes, one containing beta(wild-type) and the other Glu Tag-linked beta(E190Q), were mixed, urea-denatured, and dialyzed, and alpha(3) beta(3) gamma complexes were reconstituted. Each of the complexes containing a different number of Glu . Tag-linked beta(E190Q) was separated by anion-exchange chromatography and analyzed. The results were as follows. 1) Normal steady state ATPase activity requires three intact catalytic sites. 2) Chase-acceleration, a catalytic cooperativity, requires at least two intact catalytic sites. 3) Single-site catalysis can be mediated by a single intact catalytic site alone. Rescrambling of subunits between complexes could occur when the complex was aged under certain conditions, and this might be one of the reasons for previous contradictory results (Miwa, K., Ohtsubo, M., Denda, K., Hisabori, T., Date, T., and Yoshida, M. (1989) J. Biochem. (Tokyo) 106, 730-734).

    Web of Science

    researchmap

  • The alpha(3)beta(3)gamma complex of the F-1-ATPase from thermophilic Bacillus PS3 containing the alpha D261N substitution fails to dissociate inhibitory MgADP from a catalytic site when ATP binds to noncatalytic sites Reviewed

    JM Jault, T Matsui, FM Jault, C Kaibara, E Muneyuki, M Yoshida, Y Kagawa, WS Allison

    BIOCHEMISTRY   34 ( 50 )   16412 - 16418   1995.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER CHEMICAL SOC  

    ATP hydrolyses by the wild-type alpha(3) beta(3) gamma and mutant (alpha D261N)(3) beta(3) gamma subcomplexes of the F-1-ATPase from the thermophilic Bacillus PS3 have been compared. The wild-type complex hydrolyzes 50 mu M ATP in three kinetic phases: a burst decelerates to an intermediate phase, which then gradually accelerates to a final rate, In contrast, the mutant complex hydrolyzes 50 mu M or 2 mM ATP in two kinetic phases, The mutation abolishes acceleration from the intermediate phase to a faster final rate, Both the wild-type and mutant complexes hydrolyze 4TP with a lag after loading a catalytic site with MgADP. The rate of the MgADP-loaded wild-type complex rapidly accelerates and approaches that observed for the wild-type apo-complex. The MgADP-loaded mutant complex hydrolyzes ATP with a more pronounced lag, and the gradually accelerating rate approaches the slow, final rate observed with the mutant ape-complex. Lauryl dimethylamide oxide (LDAO) stimulates hydrolysis of 2 mM ATP catalyzed by wild-type and mutant complexes 4- and 7.5-fold, respectively. The rate of release of [H-3]ADP from the Mg[H-3]ADP-loaded mutant complex during hydrolysis of 40 mu M ATP is slower than observed with the wild-type complex. LDAO increases the rate of release of [H-3]ADP from the preloaded wild-type and mutant complexes during hydrolysis of 40 mu M ATP, Again, release is slower with the mutant complex. When the wild-type and mutant complexes are irradiated in the presence of 2-N-3-[H-3]ADP plus Mg2+ or 2-N-3-[H-3]ATP plus Mg2+ and azide, the same extent of labeling of noncatalytic sites is observed. Whereas ADP and ATP protect noncatalytic sites of the wild-type and mutant complexes about equally from labeling by 2-N-3-[H-3]ADP or 2-N-3-[H-3]ATP, respectively, AMP-PNP provides little protection of noncatalytic sites of the mutant complex. The results suggest that the substitution does not prevent binding of ADP or ATP to noncatalytic sites, but rather that it affects cross-talk between liganded noncatalytic sites and catalytic sites which is necessary to promote dissociation of inhibitory MgADP.

    Web of Science

    researchmap

  • ANALYSIS OF TIME-DEPENDENT CHANGE OF ESCHERICHIA-COLI F1-ATPASE ACTIVITY AND ITS RELATIONSHIP WITH APPARENT NEGATIVE COOPERATIVITY Reviewed

    Y KATO, T SASAYAMA, E MUNEYUKI, M YOSHIDA

    BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS   1231 ( 3 )   275 - 281   1995.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    Except for the case of gradual activation of EF(1) (F-1-ATPase from Escherichia coli) caused by the dissociation of the epsilon subunit [Laget, P.P. and Smith, J.B. (1979) Arch. Biochem. Biophys. 197, 83-89], EF(1) has long been thought not to show a time-dependent change in activity [Senior, A.E. et al. (1992) Arch. Biochem. Biophys. 297, 340-344]. Here, we report the time-dependent inactivation and activation of EF(1), which are apparently similar to those of mitochondrial F-1-ATPases [Vasilyeva, E.A. et al, (1982) Biochem. J. 202, 15-23]. Analysis of these changes as a function of ATP concentrations in relation to negative cooperativity revealed that the initial inactivation phase was attributable to the decrease in the V-max associated with the low K-m (around 10 mu M), and the following activation, probably due to the dissociation of the epsilon subunit, corresponded to the increase in the V-max associated with the high K-m (in the order of 100 mu M). Thus, the time-dependent change in EF(1) activity is closely related to the apparent negative cooperativity (multiple K-m values) of ATP hydrolysis.

    Web of Science

    researchmap

  • Direct reconstitution of bacteriorhodopsin into planar phospholipid bilayers - detergent effect Reviewed

    Mineo Ikematsu, Yukihiro Sugiyama, Masahiro Iseki, Eiro Muneyuki, Atsuo Mizukami

    Biophysical Chemistry   54 ( 2 )   155 - 164   1995

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER  

    This paper describes how the structure and concentration level of a detergent used for substitution after bacteriorhodopsin (bR) solubilization affect the reconstitution of the bR into phospholipid planar bilayers. A direct insertion method was used for the bR reconstitution into the bilayers. Two detergents representing the two major types were used: sodium deoxycholate with a cholane-ring structure, and octylglucoside with a linear (or chain) structure. We then characterized the reconstitution for the two detergents by considering the detergent separation profiles and the photocurrent variations upon addition of lanthanum chloride and the protonophore FCCP (carbonylcyanide-p-trifluoromethoxyphenylhydrazone). We found that for successful transmembrane reconstitution of bR the detergent with the cholane-ring structure was preferable to that with the linear structure when the detergent concentration was above its critical micellar concentration. This preference was explained by the ease with which the detergent with the cholane-ring structure was removed from protein compared to that with the linear structure. Finally, we proposed a scheme for the reconstitution of the protein. © 1995.

    DOI: 10.1016/0301-4622(94)00111-V

    Scopus

    researchmap

  • CATALYTIC COOPERATIVITY OF BEEF-HEART MITOCHONDRIAL F1-ATPASE REVEALED BY USING 2',3'-O-(2,4,6-TRINITROPHENYL)-ATP AS A SUBSTRATE - AN INDICATION OF MUTUALLY ACTIVATING CATALYTIC SITES Reviewed

    E MUNEYUKI, T HISABORI, WS ALLISON, JM JAULT, T SASAYAMA, M YOSHIDA

    BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS   1188 ( 1-2 )   108 - 116   1994.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    The interaction of 2',3'-O-(2,4,6-trinitrophenyl)ATP (TNP-ATP) with bovine mitochondrial F-1-ATPase (MF(1)) was examined under substoichiometric and stoichiometric conditions to investigate the relationship between the amount of bound TNP-AT(D)P and extent of inhibition on steady state ATP hydrolysis. The hydrolysis of bound TNP-ATP under substoichiometric condition proceeded slowly, with a first order rate constant of 0.014 s(-1). However, hydrolysis was greatly accelerated by addition of excess ATP. The hydrolyzed product, TNP-ADP, did not dissociate from the enzyme even after the addition of excess ATP. These properties were the same for both native and nucleotide depleted enzyme. The difference spectrum induced by binding TNP-ATP to MF(1) had a distinct peak at 410 nm and a deep trough at 395 nm, which were similar to those induced when TNP-ATP bound to the isolated beta subunit of the thermophilic F-1-ATPase. The magnitude of difference spectra as a function of TNP-ATP concentration suggested the presence of at least two types of binding sites on the MF(1) molecule. The first site, where substoichiometric TNP-ATP was hydrolyzed, had a very high affinity for TNP-ATP. TNP-AT(D)P bound to this site did not dissociate even in the presence of excess ATP. TNP-AT(D)P bound to the second site dissociated slowly when excess ATP was added. The steady state ATPase activity at 100 mu M ATP was linearly suppressed as pre-loaded TNP-ATP increased. The binding of 2 mol of TNP-ATP per mol of MF(1) was required to abolish ATPase activity. A model which assumes mutually-activating two catalytic sites is presented to explain these results.

    Web of Science

    researchmap

  • TYR-341 OF THE BETA-SUBUNIT IS A MAJOR K-M-DETERMINING RESIDUE OF TF1-ATPASE - PARALLEL EFFECT OF ITS MUTATIONS ON K-D((ATP)) OF THE BETA-SUBUNIT AND ON K-M((ATP)) OF THE ALPHA(3)BETA(3)GAMMA COMPLEX Reviewed

    M ODAKA, C KAIBARA, T AMANO, T MATSUI, E MUNEYUKI, K OGASAHARA, K YUTANI, M YOSHIDA

    JOURNAL OF BIOCHEMISTRY   115 ( 4 )   789 - 796   1994.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPANESE BIOCHEMICAL SOC  

    Residue Tyr-341 of the F-1-ATPase beta subunit from a thermophilic Bacillus strain, PS3, was mutagenized to leucine, cysteine or alanine. Each of the mutated beta subunits was isolated and its affinity for ATP-Mg was examined by means of difference circular dichroism and differential titration calorimetry. The K-d values for ATP-Mg obtained were: beta Y341 (wild type), 0.015mM; beta Y341L, 0.7mM; beta Y341C and beta Y341A, &gt;3mM. All the mutant beta subunits could be reconstituted into the alpha(3) beta(3) gamma complex with alpha and gamma subunits. The alpha(3) beta((mutant)3) gamma complexes hydrolyzed ATP with apparent V-max values larger than that of the alpha(3) beta(wild)(3) gamma complex. The apparent K-m values of the alpha(3) beta((mutant)3) gamma complexes increased in parallel with the K-d values for ATP-Mg of the isolated mutant beta subunits. These results indicate that residue beta Y341 is directly involved in the catalytic ATP-Mg binding and is a major K-m-determining residue of F-1-ATPase.

    Web of Science

    researchmap

  • COMPARISON OF THE ATPASE ACTIVITIES OF BOVINE HEART AND LIVER MITOCHONDRIAL ATP SYNTHASES WITH DIFFERENT TISSUE-SPECIFIC GAMMA-SUBUNIT ISOFORMS Reviewed

    C MATSUDA, E MUNEYUKI, H ENDO, M YOSHIDA, Y KAGAWA

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   200 ( 2 )   671 - 678   1994.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS  

    The kinetics of heart and liver mitochondrial ATPase (FoF(1)) were examined using submitochondrial particles (SMPs) purified from the two tissues to obtain information on the role of gamma subunit isoforms. The F-1 portion is mainly composed of the catalytic, common alpha beta subunits and tissue-specific gamma subunits. In contrast to the previous reports on the kinetics and crystallography of various F-1's, the Vmax and Km of the two isoforms of FoF(1) were identical although the SMPs were prepared from different tissues. Moreover sodium azide inhibited the two equally. The ATPase activity of liver SMP showed slightly steeper pH-dependency than that of heart SMP but the pH optima of the two were the same (pH 8). (C) 1994 Academic Press, Inc.

    Web of Science

    researchmap

  • Direct transmembraneous reconsitution of Bacteriorhodopsin into planar phospholipid bilayers Reviewed

    Muneyuki,E, Ikematsu,M, Iseki,M, Sugiyama,Y, Mizukami,A, Ohno,K, Yoshida,M, Hirata,H

    Biochim.Biophys.Acta   1183   171 - 179   1993.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER  

    researchmap

  • INHIBITORY EFFECT OF NAN3 ON THE F0F1 ATPASE OF SUBMITOCHONDRIAL PARTICLES AS RELATED TO NUCLEOTIDE-BINDING Reviewed

    E MUNEYUKI, M MAKINO, H KAMATA, Y KAGAWA, M YOSHIDA, H HIRATA

    BIOCHIMICA ET BIOPHYSICA ACTA   1144 ( 1 )   62 - 68   1993.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    The inhibitory effects of NaN3 on the F0F1 ATPase of beef heart submitochondrial particles were investigated. It was shown that NaN3 inhibited the ATPase activity only in the presence of ATP or ADP and the inhibition proceeded slowly. Analysis of the time-course of the inhibition process lead to a conclusion that an ATP binding site which has an apparent K(d) of 14.0 +/- 8.7 muM is responsible for the increase of NaN3 sensitivity. This value agreed well with the low K(m) of ATP hydrolysis characterized before (Muneyuki, E., and Hirata, H. (1988) FEBS Lett. 234, 455-458) and in the range of so-called bi-site catalysis. The same conclusion was derived as for isolated F1 ATPase. From similar analysis, the K(d) of this site for ADP was deduced to be 1.34 +/- 0.45 muM, which also agreed with that reported by Pedersen (Pedersen, P.L. (1975) Biochem. Biophys. Res. Commun. 64, 610-616) and also in the same range as reported for the low K(m) of ATP synthesis by activated submitochondrial particles. These results suggest that hydrolysis through the low K(m) mode of ATPase reaction leads the enzyme NaN3 sensitive form and this reaction cycle corresponds to the low K(m) mode of ATP synthesis.

    Web of Science

    researchmap

  • A chaperonin from a thermophilic bacterium, Thermus thermophilus. Reviewed

    M. Yoshida, N. Ishii, E. Muneyuki, H. Taguchi

    Philosophical transactions of the Royal Society of London. Series B, Biological sciences   339 ( 1289 )   305 - 312   1993.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:The Royal Society(UK,London)  

    Unlike Escherichia coli chaperonins, a chaperonin (cpn) from a thermophilic bacterium, Thermus thermophilus, consisting of homologues to GroEL (cpn 60) and GroES (cpn 10) is co-purified as a large complex. Thermus chaperonin shows a bullet-like shape in the side view seen by electron microscopy, and antibody against cpn 10 binds only to the round side of the bullet. We conclude that a single cpn 60-heptamer ring with two stripes stacks into two layers and a cpn 10 oligomer binds to one side of the layers. The purified Thermus chaperonin contains endogenously bound ADP, and incubation with ATP causes a partial dissociation of chaperonin into cpn 60 monomers and a cpn 10 heptamer. The effect of Thermus chaperonin on protein refolding upon dilution from guanidine HC1 is different at three temperature ranges. At high temperatures above 55 degrees C, where the native proteins are stable but their spontaneous foldings fail, the chaperonin induces productive folding in an ATP-dependent manner. At middle temperatures (25-55 degrees C) where spontaneous foldings of the enzymes occur, the chaperonin slows down the rate of folding without changing the final yield of productive folding. At lower temperatures below 25 degrees C where spontaneous foldings also occur, the chaperonin arrests the folding even in the presence of ATP. When a solution of relatively heat labile protein is incubated at high temperatures, and then residual activity of the protein is measured at its optimal temperature after incubation with ATP, the temperature that causes irreversible heat denaturation of the protein is elevated about 10 degrees C by inclusion of Thermus chaperonin in the solution.(ABSTRACT TRUNCATED AT 250 WORDS)

    DOI: 10.1098/rstb.1993.0029

    Scopus

    PubMed

    researchmap

  • SINGLE SITE HYDROLYSIS OF 2',3'-O-(2,4,6-TRINITROPHENYL)-ATP BY THE F1-ATPASE FROM THE THERMOPHYLIC BACTERIUM PS3 IS ACCELERATED BY THE CHASE-ADDITION OF EXCESS ATP Reviewed

    T HISABORI, E MUNEYUKI, M ODAKA, K YOKOYAMA, K MOCHIZUKI, M YOSHIDA

    JOURNAL OF BIOLOGICAL CHEMISTRY   267 ( 7 )   4551 - 4556   1992.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    The interaction of 2',3'-O-(2,4,6-trinitrophenyl)adenosine 5'-triphosphate (TNP-ATP) and TNP-ADP to F1-ATPase from a thermophilic bacterium PS3 (TF1) was investigated. When TNP-ADP or TNP-ATP was added to the isolated alpha or beta-subunit of TF1, characteristic difference spectra were generated for each subunit. Difference spectra generated on addition of these analogs to TF1 resembled those observed for the beta-subunit, indicating TNP analogs bind to the beta-subunits in the molecule of TF1. Results of equilibrium dialysis showed that TNP-ADP binds to a single high affinity site on TF1 in the presence of Mg2+ with a dissociation constant of 2.2 nM. When TNP-ATP was added to TF1 in a substoichiometric molar ratio, it rapidly bound to TF1 and was slowly hydrolyzed. The hydrolysis proceeded nearly to completion without showing stable equilibrium between bound species of TNP-ATP and TNP-ADP. Similar to beef heart mitochondrial F1, this hydrolysis was greatly accelerated by the chase-addition of 100-mu-M ATP. However, the hydrolyzed product, TNP-ADP, remained bound on the beta-subunit even after the chase.

    Web of Science

    researchmap

  • HETEROGENEOUS HYDROLYSIS OF SUBSTOICHIOMETRIC ATP BY THE F1-ATPASE FROM ESCHERICHIA-COLI Reviewed

    E MUNEYUKI, M YOSHIDA, DA BULLOUGH, WS ALLISON

    BIOCHIMICA ET BIOPHYSICA ACTA   1058 ( 2 )   304 - 311   1991.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    The hydrolysis of 0.3-mu-M [alpha, gamma-P-32]ATP by 1-mu-M F1-ATPase isolated from the plasma membranes of Escherichia coli has been examined in the presence and absence of inorganic phosphate. The rate of binding of substoichiometric substrate to the ATPase is attenuated by 2 mM phosphate and further attenuated by 50 mM phosphate. Under all conditions examined, only 10-20% of the [alpha, gamma-P-32]ATP that bound to the enzyme was hydrolyzed sufficiently slowly to be examined in cold chase experiments with physiological concentrations of non-radioactive ATP. These features differ from those observed with the mitochondrial F1-ATPase. The amount of bound substrate in equilibrium with bound products observed in the slow phase which was subject to promoted hydrolysis by excess ATP was not affected by the presence of phosphate. Comparison of the fluxes of enzyme-bound species detected experimentally in the presence of 2 mM phosphate with those predicted by computer simulation of published rate constants determined for uni-site catalysis (Al-Shawi, M.D., Parsonage, D. and Senior, A.E. (1989) J. Biol. Chem. 264, 15376-15383) showed that hydrolysis of substoichiometric ATP observed experimentally was clearly biphasic. Less than 20% of the substoichiometric ATP added to the enzyme was hydrolyzed according to the published rate constants which were calculated from the slow phase of product release in the presence of 1 mM phosphate. The majority of the substoichiometric ATP added to the enzyme was hydrolyzed with product release that was too rapid to be detected by the methods employed in this study, indicating again that the F1-ATPase from E. coli and bovine heart mitochondria hydrolyze substoichiometric ATP differently.

    Web of Science

    researchmap

  • AROMATIC RINGS OF TYROSINE RESIDUES AT ADENINE-NUCLEOTIDE BINDING-SITES OF THE BETA-SUBUNITS OF F1-ATPASE ARE NOT NECESSARY FOR ATPASE ACTIVITY Reviewed

    M ODAKA, H KOBAYASHI, E MUNEYUKI, M YOSHIDA

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   168 ( 1 )   372 - 378   1990.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Web of Science

    researchmap

  • STEADY-STATE KINETICS OF PROTON TRANSLOCATION CATALYZED BY THERMOPHILIC F0F1-ATPASE RECONSTITUTED IN PLANAR BILAYER-MEMBRANES Reviewed

    E MUNEYUKI, Y KAGAWA, H HIRATA

    JOURNAL OF BIOLOGICAL CHEMISTRY   264 ( 11 )   6092 - 6096   1989.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Web of Science

    researchmap

  • KINETIC-ANALYSIS OF PROTON TRANSLOCATION CATALYZED BY F0F1 ATPASE Reviewed

    E MUNEYUKI, H HIRATA

    FEBS LETTERS   234 ( 2 )   455 - 458   1988.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    Web of Science

    researchmap

  • RECONSTITUTION OF THE PROTON TRANSLOCATING ATPASE FROM BOVINE HEART-MITOCHONDRIA INTO PLANAR PHOSPHOLIPID-BILAYER MEMBRANES Reviewed

    E MUNEYUKI, K OHNO, Y KAGAWA, H HIRATA

    JOURNAL OF BIOCHEMISTRY   102 ( 6 )   1433 - 1440   1987.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPANESE BIOCHEMICAL SOC  

    Web of Science

    researchmap

  • AN ACTIN-DEPOLYMERIZING PROTEIN (DESTRIN) FROM PORCINE KIDNEY - ITS ACTION ON F-ACTIN CONTAINING OR LACKING TROPOMYOSIN Reviewed

    E NISHIDA, E MUNEYUKI, S MAEKAWA, Y OHTA, H SAKAI

    BIOCHEMISTRY   24 ( 23 )   6624 - 6630   1985

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER CHEMICAL SOC  

    Web of Science

    researchmap

  • PURIFICATION OF COFILIN, A 21,000 MOLECULAR-WEIGHT ACTIN-BINDING PROTEIN, FROM PORCINE KIDNEY AND IDENTIFICATION OF THE COFILIN-BINDING SITE IN THE ACTIN SEQUENCE Reviewed

    E MUNEYUKI, E NISHIDA, K SUTOH, H SAKAI

    JOURNAL OF BIOCHEMISTRY   97 ( 2 )   563 - 568   1985

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPANESE BIOCHEMICAL SOC  

    Web of Science

    researchmap

  • ACTION OF A 19K-PROTEIN FROM PORCINE BRAIN ON ACTIN POLYMERIZATION - A NEW FUNCTIONAL CLASS OF ACTIN-BINDING PROTEINS Reviewed

    E NISHIDA, S MAEKAWA, E MUNEYUKI, H SAKAI

    JOURNAL OF BIOCHEMISTRY   95 ( 2 )   387 - 398   1984

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:JAPANESE BIOCHEMICAL SOC  

    Web of Science

    researchmap

▼display all

Books

  • 化学フロンティアシリーズ 23 1分子ナノバイオ計測

    鳥谷部祥一, 宗行英朗, 編者, 野地博行(第10章 分子モーターの1分子実験熱力学)

    化学同人  2014 

     More details

    Responsible for pages:131-138   Language:Japanese  

    researchmap

  • DOJIN BIOSCIENCE SERIES 17 一分子生物学

    上野博史, 鳥谷部祥一, 宗行英朗, 編者, 原田 慶恵, 石渡, 信一(第7章FoF1モーター)

    化学同人  2014 

     More details

    Responsible for pages:88-101   Language:Japanese  

    researchmap

  • 中央大学学術シンポジウム研究叢書 9 ナノスケール・ミクロスケールから見える ビックな世界 (新藤 斎 編著 )

    鳥谷部祥一, 宗行英朗( Role: Joint author第一章 マクスウェルの悪魔の実現|rn| ―― ミクロの世界で情報を使って熱エネルギーを自由エネルギーに変換する ――)

    中央大学出版部  2013 

     More details

    Total pages:19   Responsible for pages:19   Language:Japanese   Book type:Scholarly book

    researchmap

  • 石渡信一,桂勲,桐野豊,美宅成樹編, 生物物理学ハンドブック, 朝倉書店, 東京, 2007, xv+662p, 26.5×19cm, 本体28,000円[大学院向], ISBN978-4-254-17122-8

    宗行 英朗

    一般社団法人日本物理学会  2008.2 

     More details

    Language:Japanese  

    researchmap

  • Handbook of ATPases, Capter 10 Rotation of F1-ATPase(Eiro Muneyuki and Masasuke Yoshida)

    Eiro Muneyuki, Masasuke Yoshida, ed. M.Futai, Y.Wada, J.Kaplan( Role: Sole authorCapter 10 Rotation of F1-ATPase)

    WILEY-VCH Verlag GmbH & Co. Weinheim  2004.8 

     More details

    Responsible for pages:261-287   Language:English   Book type:Scholarly book

    researchmap

  • 光合成辞典

    日本光合成研究会編( Role: Sole author「回転触媒説」ほか7項目)

    学会出版センター  2003.11 

     More details

    Total pages:430   Language:Japanese   Book type:Scholarly book

    researchmap

  • シリーズ・ニューバイオフィジックスⅡ-10 生物物理学とはなにか

    宗行英朗, 吉田賢右, 編者, 曽我部正博, 郷信広( Role: Sole author第1章7.エネルギー転換:ATP合成酵素)

    共立出版  2003.9 

     More details

    Responsible for pages:88-100   Language:Japanese   Book type:Scholarly book

    researchmap

  • 新ミトコンドリア学

    宗行英朗, 吉田賢右, 編者, 内海耕造, 井上正康( Role: Sole author第一章1.3エネルギー転換系と分子モーター)

    共立出版  2001.11 

     More details

    Responsible for pages:14-21   Language:Japanese   Book type:Scholarly book

    researchmap

  • シリーズ・ニューバイオフィジックスⅡ-3 ポンプとトランスポーター

    久堀徹, 宗行英朗, 野地博行, 吉田賢右, 編者, 平田肇, 茂木立志( Role: Sole author第1章1. ATP合成酵素)

    共立出版  2000.6 

     More details

    Responsible for pages:19-40   Language:Japanese   Book type:Scholarly book

    researchmap

▼display all

MISC

  • Thermodynamic role of main reaction pathway and multi-body information flow in membrane transport

    Satoshi Yoshida, Yasushi Okada, Eiro Muneyuki, Sosuke Ito

    2021.12

     More details

    The two classes of membrane transport, namely, secondary active and passive
    transport, are understood as different reaction pathways in the same protein
    structure, modeled by the 16-state model in this paper. To quantify the
    thermodynamic difference between secondary active transport and passive
    transport, we extend the second law of information thermodynamics of the
    autonomous demon in the four-state model composed of two subsystems to the
    16-state model composed of four subsystems representing the membrane transport.
    We reduce the 16 states to 4 states and derive the coarse-grained second law of
    information thermodynamics, which provides an upper bound of the free energy
    transport by the coarse-grained information flow. We also derive an upper bound
    on the free energy transport by the multi-body information flow representing
    the two-body or four-body correlations in the 16-state model by exploiting the
    cycle decomposition. The coarse-grained information flow and the multi-body
    information flows express the quantitative difference between secondary active
    and passive transport. The numerical analysis shows that the coarse-grained
    information flow is positive for secondary active transport and negative for
    passive transport. The four-body correlation is dominant in the multi-body
    information flows for secondary active transport. In contrast, the two-body
    correlation is dominant for passive transport. This result shows that both the
    sign of the coarse-grained information flow and the difference of the four-body
    correlation and the two-body correlation explain the difference of the free
    energy transport in secondary active and passive transport.

    arXiv

    researchmap

    Other Link: http://arxiv.org/pdf/2112.04024v1

  • F1‐ATPaseのP‐loop変異体TF1(βG158A)のリン酸解離角度の解明

    成田宏夏, 星名仁志, 吉田光, 中山洋平, 鳥谷部祥一, 上野博史, 宗行英朗

    日本物理学会講演概要集(CD-ROM)   71 ( 2 )   ROMBUNNO.14pBE‐9 - 3051   2016.9

     More details

    Language:Japanese   Publisher:一般社団法人 日本物理学会  

    &lt;p&gt;ATP合成酵素であるF1-ATPaseの反応スキームについて生成物のADPとPiの解離の関係におけるモデルが2つ考えられている。2010年に渡邊らはADPよりもPiが先に解離することを報告したが、下らはADPはPiよりも後に解離することを報告した。そこで、本研究で、私たちは変異体を用いてこの問題を議論することにした。P-loop変異体TF1(βG158A)を用いた一分子回転観察実験を行い、ATP加水分解とPi解離が遅いことを確認した。更に、ハイブリッドF1変異体を作成し、同様に回転実験から得られたdwell timeの解析を行うことで、PiはADPよりも先に解離するモデルを示唆する結果を得た。&lt;/p&gt;

    DOI: 10.11316/jpsgaiyo.71.2.0_3051

    J-GLOBAL

    researchmap

  • 28pBD-6 Biological molecular motor and fluctuation

    Muneyuki Eiro

    Meeting abstracts of the Physical Society of Japan   69 ( 1 )   340 - 340   2014.3

     More details

    Language:Japanese   Publisher:The Physical Society of Japan (JPS)  

    CiNii Books

    researchmap

  • 1P163 The response of TF1 βE190D mutant to the external torque(11. Molecular motor,Poster,The 52nd Annual Meeting of the Biophysical Society of Japan(BSJ2014))

    Tanaka Mana, Kawakami Tomohiro, Toyabe Shoichi, Ueno Hiroshi, Kudo Seishi, Muneyuki Eiro

    Seibutsu Butsuri   54 ( 1 )   S168   2014

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.54.S168_1

    researchmap

  • 3P152 Nucleotide binding to TF1 β subunit in relation to the effect of Pi(Molecular motor,Poster,The 52th Annual Meeting of the Biophysical Society of Japan(BSJ2014))

    Riku Nagano, Kiyoshi Obara, Hiroshi Ueno, Eiro Muneyuki

    Seibutsu Butsuri   54 ( 1 )   S274   2014

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.54.S274_2

    researchmap

  • 1P161 Observation of the rotation of F1-ATPase under the constant external torque(11. Molecular motor,Poster,The 52nd Annual Meeting of the Biophysical Society of Japan(BSJ2014))

    Kikuchi Yohsuke, Ueno Hiroshi, Nakayama Takahiro, Muneyuki Eiro, Toyabe Shouichi

    Seibutsu Butsuri   54 ( 1 )   S167   2014

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.54.S167_5

    researchmap

  • 1P160 Inhibitory effect of Pi on F_1-ATPase P-loop mutant TF_1(βG158A)(11. Molecular motor,Poster,The 52nd Annual Meeting of the Biophysical Society of Japan(BSJ2014))

    Hoshina Hitoshi, Yoshida Hikaru, Ito Ayumi, Ito Jotaro, Toyabe Shoichi, Ueno Hiroshi, Muneyuki Eiro

    Seibutsu Butsuri   54 ( 1 )   S167   2014

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.54.S167_4

    researchmap

  • 1P174 Direct observation of the rotation of V_1-ATPase from Enterococcus hirae and its torque(11.Molecular motor,Poster,The 51st Annual Meeting of the Biophysical Society of Japan)

    Ueno Hiroshi, Minagawa Yoshihiro, Yamato Ichiro, Murata Takeshi, Iino Ryota, Muneyki Eiro

    Seibutsu Butsuri   53 ( 1 )   S134   2013

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.53.S134_5

    researchmap

  • 1P175 The relationship between F_1-ATPase P-loop mutants and Pi release(11.Molecular motor,Poster,The 51st Annual Meeting of the Biophysical Society of Japan)

    Yoshida Hikaru, Ito Ayumi, Ito Jotaro, Masaike Tomoko, Nishizaka Takayuki, Toyabe Shoichi, Ueno Hiroshi, Muneyuki Eiro

    Seibutsu Butsuri   53 ( 1 )   S134   2013

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.53.S134_6

    researchmap

  • Experimental thermodynamics of single molecular motor.

    Shoichi Toyabe, Eiro Muneyuki

    Biophysics (Nagoya-shi, Japan)   9   91 - 8   2013

     More details

    Language:English   Publishing type:Article, review, commentary, editorial, etc. (scientific journal)  

    Molecular motor is a nano-sized chemical engine that converts chemical free energy to mechanical motions. Hence, the energetics is as important as kinetics in order to understand its operation principle. We review experiments to evaluate the thermodynamic properties of a rotational F1-ATPase motor (F1-motor) at a single-molecule level. We show that the F1-motor achieves 100% thermo dynamic efficiency at the stalled state. Furthermore, the motor reduces the internal irreversible heat inside the motor to almost zero and achieves a highly-efficient free energy transduction close to 100% during rotations far from quasistatic process. We discuss the mechanism of how the F1-motor achieves such a high efficiency, which highlights the remarkable property of the nano-sized engine F1-motor.

    DOI: 10.2142/biophysics.9.91

    PubMed

    researchmap

  • Information-to-free-energy conversion: Utilizing thermal fluctuations.

    Shoichi Toyabe, Eiro Muneyuki

    Biophysics (Nagoya-shi, Japan)   9   107 - 12   2013

     More details

    Language:English   Publishing type:Article, review, commentary, editorial, etc. (scientific journal)  

    Maxwell's demon is a hypothetical creature that can convert information to free energy. A debate that has lasted for more than 100 years has revealed that the demon's operation does not contradict the laws of thermodynamics; hence, the demon can be realized physically. We briefly review the first experimental demonstration of Maxwell's demon of Szilard's engine type that converts information to free energy. We pump heat from an isothermal environment by using the information about the thermal fluctuations of a Brownian particle and increase the particle's free energy.

    DOI: 10.2142/biophysics.9.107

    PubMed

    researchmap

  • 1P177 Comparison of the nucleotide binding to the isolated βsubunit and the F1-ATPase using the Sopped-Flow method(11.Molecular motor,Poster,The 51st Annual Meeting of the Biophysical Society of Japan)

    Nagano Riku, Obara Kiyoshi, Masaike Tomoko, Ueno Hiroshi, Muneyuki Eiro

    Seibutsu Butsuri   53 ( 1 )   S135   2013

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.53.S135_2

    researchmap

  • 熱ゆらぎを利用する―情報熱機関の実現―

    鳥谷部 祥一, 宗行 英朗

    生物物理   52 ( 3 )   136 - 139   2012.5

     More details

    Language:Japanese   Publishing type:Article, review, commentary, editorial, etc. (scientific journal)  

    researchmap

  • 27aPS-27 Recognition Mechanism of Presequence by Mitochondrial Outer Membrane Protein Tom20 using Replica-exchange Molecular Dynamics Simulation

    Komuro Yasuaki, Mori Takaharu, Miyashita Naoyuki, Muneyuki Eiro, Saitoh Takashi, Kohda Daisuke, Sugita Yuji

    Meeting abstracts of the Physical Society of Japan   67 ( 1 )   429 - 429   2012.3

     More details

    Language:Japanese   Publisher:The Physical Society of Japan (JPS)  

    CiNii Books

    researchmap

  • 1PS024 Inhibitory effect of Pi on F_1-ATPase P-loop mutant(The 50th Annual Meeting of the Biophysical Society of Japan)

    Yoshida Hikaru, Ito Ayumi, Ito Jotaro, Toyabe Syoichi, Ueno Hiroshi, Muneyuki Eiro

    Seibutsu Butsuri   52 ( 0 )   S78   2012

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.52.S78_2

    researchmap

  • 2SI-02 Single molecule energetics of F_1-ATPase motor(2SI The art of energetic and functional efficiency in biomoiecular machines on the single molecule level,Symposium,The 50th Annual Meeting of the Biophysical Society of Japan)

    Toyabe Shoichi, Watanabe-Nakayama Takahiro, Ueno Hiroshi, Okamoto Tetsuaki, Taketani Hiroshi, Kudo Seishi, Muneyuki Eiro

    Seibutsu Butsuri   52 ( 0 )   S17   2012

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.52.S17_3

    researchmap

  • 1PS032 Simultaneous observation of fluorescent-nucleotides and rotation of rotary molecular motor on plasmonic metal nanostructures(The 50th Annual Meeting of the Biophysical Society of Japan)

    Ueno Hiroshi, Toyabe Shoichi, Muneyuki Eiro

    Seibutsu Butsuri   52 ( 0 )   S79   2012

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.52.S79_4

    researchmap

  • 1L1348 Simultaneous observation of fluorescent-nucleotides and rotation of F1-ATPase using plasmonic gold nanoparticle(Molecular motor 1,The 49th Annual Meeting of the Biophysical Society of Japan)

    Ueno Hiroshi, Toyabe Shoichi, Muneyuki Eiro

    Seibutsu Butsuri   51 ( 0 )   S60   2011

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.51.S60_4

    researchmap

  • 金微粒子によるプラズモン増強電場を利用した蛍光ATPとF1‐ATPaseの回転の同時観察

    上野博史, 鳥谷部祥一, 宗行英朗

    日本生体エネルギー研究会討論会講演要旨集   37th   56 - 57   2011

     More details

    Language:Japanese  

    J-GLOBAL

    researchmap

  • 3A1422 pH dependency of photoelectric current of bacteriorhodopsin absorbed on Lumirror membrane(3A Biol & Artifi memb 4: Transport, Signal transduction,The 49th Annual Meeting of the Biophysical Society of Japan)

    Miyazaki Shiduma, Toyabe Shoichi, Ueno Hiroshi, Muneyuki Eiro

    Seibutsu Butsuri   51 ( 0 )   S107   2011

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.51.S107_1

    researchmap

  • 1L1548 P05 1YE1000 Single-molecule nonequilibrium thermodynamics of molecular motor(Molecular motor 1,Early Research in Biophysics Award Candidate Presentations,Early Research in Biophysics Award,The 49th Annual Meeting of the Biophysical Society of Japan

    Toyabe Shoichi, Ueno Hiroshi, Muneyuki Eiro

    Seibutsu Butsuri   51 ( 0 )   S62   2011

     More details

    Language:English   Publisher:一般社団法人 日本生物物理学会  

    DOI: 10.2142/biophys.51.S62_2

    researchmap

  • 26aTF-9 Estimating potential profile of molecular motor from single-molecule trajectory

    Toyabe Shoichi, Ueno Hiroshi, Muneyuki Eiro

    Meeting Abstracts of the Physical Society of Japan   66 ( 0 )   308 - 308   2011

     More details

    Language:Japanese   Publisher:一般社団法人 日本物理学会  

    CiNii Books

    researchmap

  • Single-molecule nonequilibrium energetics of a molecular-motor F1-ATPase

    物性研究   95 ( 3 )   320 - 322   2010.12

     More details

    Language:Japanese   Publishing type:Article, review, commentary, editorial, etc. (scientific journal)  

    CiNii Books

    researchmap

    Other Link: http://dl.ndl.go.jp/info:ndljp/pid/10942999

  • 23aTF-3 Converting information to energy by feedback control

    Toyabe Shoichi, Sagawa Takahiro, Ueda Masahito, Muneyuki Eiro, Sano Masaki

    Meeting abstracts of the Physical Society of Japan   65 ( 2 )   212 - 212   2010.8

     More details

    Language:Japanese   Publisher:The Physical Society of Japan (JPS)  

    CiNii Books

    researchmap

  • Nonequilibrium Energetics of a Single F-1-ATPase Molecule

    Shoichi Toyabe, Takahiro Watanabe-Nakayama, Tetsuaki Okamoto, Seishi Kudo, Eiro Muneyuki

    BIOPHYSICAL JOURNAL   98 ( 3 )   167A - 168A   2010.1

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:CELL PRESS  

    Web of Science

    researchmap

  • 高時間・空間分解能を持つシンプルな暗視野顕微鏡の蛍光同時観察への応用

    上野博史, 飯野亮太, 田端和仁, 榊原昇一, 宗行英朗, 野地博行

    日本生体エネルギー研究会討論会講演要旨集   36th   2010

  • 3P169 Thermodynamic parameters of nucleotide binding to the catalytic sites of Fl-ATPase revealed by fluorescence measurement.(Molecular motor,The 48th Annual Meeting of the Biophysical Society of Japan)

    Kikuchi Yosuke, Naka Yusuke, Osakabe Hidemitsu, Toyabe Shoichi, Ueno Hiroshi, Muneyuki Eiro

    Seibutsu Butsuri   50 ( 2 )   S174   2010

     More details

    Language:English   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.50.S174_5

    researchmap

  • レーザ光による金表面でのナノ物質操作=DNAの伸張固定およびナノ微粒子の収集を中心に=

    藤井翔, 芳賀正明, 宗行英朗

    光アライアンス   21 ( 12 )   19 - 23   2010

     More details

    Language:Japanese   Publishing type:Article, review, commentary, editorial, etc. (scientific journal)  

    CiNii Books

    researchmap

  • 27pQE-6 Stall torque of F_1-ATPase

    Toyabe Shoichi, Watanabe-Nakayama Takahiro, Okamoto Tetsuaki, Kudo Seishi, Muneyuki Eiro

    Meeting Abstracts of the Physical Society of Japan   64 ( 0 )   287 - 287   2009

     More details

    Language:Japanese   Publisher:一般社団法人 日本物理学会  

    CiNii Books

    researchmap

  • 23pYD-11 Molecular dynamics study of ε subunit of F_1-ATPase

    Muneyuki Eiro, Tsuji Yosuke, Sugita Yuji

    Meeting abstracts of the Physical Society of Japan   63 ( 1 )   359 - 359   2008.2

     More details

    Language:Japanese   Publisher:The Physical Society of Japan (JPS)  

    CiNii Books

    researchmap

  • 外部トルク存在下でのF1モーターの回転(Rotation of F1-motor in the presence of external torque)

    中山 隆宏, 渡部, 吉田 賢右, 杉山 滋, 工藤 成史, 宗行 英朗

    生物物理   47 ( Suppl.1 )   S242 - S242   2007.11

     More details

    Language:English   Publisher:(一社)日本生物物理学会  

    researchmap

  • F1-ATPase rotates by an asymmetric, sequential mechanism using all three catalytic subunits(vol 14, pg 841, 2007)

    Takayuki Ariga, Eiro Muneyuki, Masasuke Yoshida

    NATURE STRUCTURAL & MOLECULAR BIOLOGY   14 ( 10 )   984 - 984   2007.10

     More details

    Language:English   Publisher:NATURE PUBLISHING GROUP  

    Web of Science

    researchmap

  • 18pTC-9 A model of autonomous free energy transducer based on coupled ratchet subsystems.

    Muneyuki Eiro, Sekimoto Ken

    Meeting abstracts of the Physical Society of Japan   62 ( 1 )   331 - 331   2007.2

     More details

    Language:Japanese   Publisher:The Physical Society of Japan (JPS)  

    CiNii Books

    researchmap

  • 3P158 Rotation of F_1-motor in the presence of external torque(Molecular motors,Poster Presentations)

    Nakayama-Watanabe Takahiro, Yoshida Masasuke, Sugiyama Shigeru, Kudo Seishi, Muneyuki Eiro

    Seibutsu Butsuri   47 ( 0 )   S242   2007

     More details

    Language:English   Publisher:一般社団法人 日本生物物理学会  

    DOI: 10.2142/biophys.47.S242_3

    researchmap

  • 生体膜における水素イオン濃度勾配の利用

    宗行 英朗

    化学と教育   55 ( 12 )   610 - 613   2007

     More details

    Language:Japanese   Publishing type:Article, review, commentary, editorial, etc. (scientific journal)  

    DOI: 10.20665/kakyoshi.55.12_610

    CiNii Books

    researchmap

  • 26pYW-7 Direct observation of rotation with interim state in Fi-ATPase

    Tsumuraya M., Oiwa K., Muneyuki E., Nishizaka T.

    Meeting abstracts of the Physical Society of Japan   60 ( 1 )   367 - 367   2005.3

     More details

    Language:Japanese   Publisher:The Physical Society of Japan (JPS)  

    CiNii Books

    researchmap

  • 2P176 Application of external force with traveling wave electric field to F_1 motor

    Watanabe-Nakayama T, Muneyuki E, Sugiyama S, Kudo S, Yoshida M

    Seibutsu Butsuri   45 ( 0 )   S163   2005

     More details

    Language:Japanese   Publisher:一般社団法人 日本生物物理学会  

    DOI: 10.2142/biophys.45.S163_4

    researchmap

  • 14aPS-85 Site-Occupancy and ATPase Activity of F_1 ATPase (ΔNC mutant)

    Kon Rieko, Muneyuki Eiro, Sakaki Naoyoshi, Adachi Kengo, Yoshida Masasuke, Kinosita Kazuhiko Jr

    Meeting abstracts of the Physical Society of Japan   59 ( 2 )   258 - 258   2004.8

     More details

    Language:Japanese   Publisher:The Physical Society of Japan (JPS)  

    CiNii Books

    researchmap

  • "Chemomechanical coupling in F1-ATPase revealed by simultaneous observation of nucleotide kinetics and rotation"

    Takayuki Nishizaka, Kazuhiro Oiwa, Hiroyuki Noji, Shigeki Kimura, Eiro Muneyuki, Masasuke Yoshida, Kazuhiko Kinosita, J

    Nature Structural & Molecular biology   11 ( 2 )   142 - 148   2004

  • Analysis of the two catalytic events that induce a 30 degrees substep rotation of F-1-ATPase

    K Shimabukuro, E Muneyuki, M Yoshida

    BIOPHYSICAL JOURNAL   84 ( 2 )   456A - 457A   2003.2

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:BIOPHYSICAL SOCIETY  

    Web of Science

    researchmap

  • 2O1345 F_1-ATPase Changes its COnformations upon Phosphate Release

    Masaike T, Muneyuki E, Noji H, Kinosita K, Yoshida M

    Seibutsu Butsuri   42 ( 2 )   S144   2002

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    DOI: 10.2142/biophys.42.S144_4

    researchmap

  • Effect of free energy of ATP hydrolysis on the rotation of rsubunit in F1-ATPase

    Noji H., Yasuda R., Muneyuki E., Yoshida M., Kinoshita K. Jr

    Biophysics   38 ( 2 )   S43   1998.9

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    CiNii Books

    researchmap

  • Transport properties of halorhodopsin

    Muneyuki E., Okuno D., Asaumi M., Yoshida M., Mogi T.

    Biophysics   38 ( 2 )   S126   1998.9

     More details

    Language:Japanese   Publisher:The Biophysical Society of Japan General Incorporated Association  

    CiNii Books

    researchmap

  • バクテリオロドプシンによる光プロトンポンプの輸送特性とイオンポンプの確率的モデル-イオンポンプとイオンチャンネルの接点-

    宗行英朗

    膜   22 ( 6 )   322 - 330   1997.6

     More details

    Language:Japanese   Publishing type:Article, review, commentary, editorial, etc. (other)   Publisher:日本膜学会  

    researchmap

  • Delta mu H+ dependency of proton translocation by bacteriorhodopsin and a stochastic energization-relaxation channel model (vol 100, pg 19688, 1996)

    E Muneyuki, M Ikematsu, M Yoshida

    JOURNAL OF PHYSICAL CHEMISTRY B   101 ( 5 )   846 - 846   1997.1

     More details

    Language:English   Publisher:AMER CHEMICAL SOC  

    Web of Science

    researchmap

  • 地上最小のモーター,ATP合成酵素 Invited

    吉田 賢右, 野地 博行, 宗行 英朗

    蛋白質 核酸 酵素   42 ( 9 )   1396 - 1406   1997

     More details

    Language:Japanese   Publishing type:Article, review, commentary, editorial, etc. (trade magazine, newspaper, online media)   Publisher:共立出版  

    researchmap

  • 地上最小のモーター,ATP合成酵素 Invited

    吉田 賢右, 野地 博行, 宗行 英朗

    蛋白質 核酸 酵素   42 ( 9 )   1396 - 1406   1997

     More details

    Language:Japanese   Publishing type:Article, review, commentary, editorial, etc. (trade magazine, newspaper, online media)   Publisher:共立出版  

    CiNii Books

    researchmap

  • 触媒部位の微小変化とサブユニットの全体変化

    天野 豊己, 柳沢 諭, 野地 博行, 宗行 英朗, 久堀 徹, 吉田 賢右

    日本分子生物学会年会プログラム・講演要旨集   19   266 - 266   1996.8

     More details

    Language:Japanese  

    CiNii Books

    researchmap

  • バクテリオロドプシンによるプロトン輸送の△pH及び△Ψ依存性

    宗行 英朗, 吉田 賢右

    日本分子生物学会年会プログラム・講演要旨集   19   271 - 271   1996.8

     More details

    Language:Japanese  

    CiNii Books

    researchmap

  • F_1-ATPase分子内でのβサブユニット同士の接触

    野地 博行, 天野 豊己, 宗行 英朗, 久堀 徹, 吉田 賢右

    日本分子生物学会年会プログラム・講演要旨集   19   267 - 267   1996.8

     More details

    Language:Japanese  

    CiNii Books

    researchmap

  • 好熱菌 F1-ATPaseにおけるヌクレオチドの結合と加水分解活性の解析

    平尾 潤, 宗行 英朗, 吉田 賢右

    日本分子生物学会年会プログラム・講演要旨集   19   266 - 266   1996.8

     More details

    Language:Japanese  

    CiNii Books

    researchmap

  • 平面リン脂質二重層膜を使ったイオンチャンネルの測定

    浜本敏郎, 宗行英朗, 平田肇

    細胞工学   7 ( 1 )   69 - 78   1988.1

     More details

    Language:Japanese   Publishing type:Article, review, commentary, editorial, etc. (other)   Publisher:秀潤社  

    researchmap

  • 平面リン脂質二重層膜を使ったイオンチャネルの測定

    浜本 敏郎, 宗行 英朗, 平田 肇

    細胞工学   7 ( 1 )   69 - 78   1988

     More details

    Language:Japanese   Publishing type:Article, review, commentary, editorial, etc. (trade magazine, newspaper, online media)  

    CiNii Books

    researchmap

▼display all

Presentations

▼display all

Research Projects

  • Understanding molecular machines from the viewpoint of information thermodynamics

    Grant number:22H04852  2022.4 - 2024.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)  Chuo University

      More details

    Grant amount: \3900000 ( Direct Cost: \3000000 、 Indirect Cost: \900000 )

    researchmap

  • Understanding the energy transduction by F1 motor from the view point of information thermodynamics

    Grant number:20H05545  2020.4 - 2022.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)  Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)  Chuo University

    Eiro, Muneyuki

      More details

    Grant amount: \2860000 ( Direct Cost: \2200000 、 Indirect Cost: \660000 )

    researchmap

  • F1モーターが準静的に動作できる限界を探る.

    2018.4 - 2021.3

    Chuo University  Grant-in-Aid for Scientific Research (C) 

    宗行 英朗

      More details

    Authorship:Principal investigator  Grant type:Competitive

    Grant amount: \4420000 ( Direct Cost: \3400000 、 Indirect Cost: \1020000 )

    researchmap

  • ATPのエネルギー総括研究

    2008.4 - 2009.3

    文部科学省  科学研究費補助金(日本学術振興会・文部科学省)-新学術領域研究(研究領域提案型) 

    鈴木 誠

      More details

    Grant type:Competitive

    「水を主役としたATPエネルギー変換」を解明するための領域研究が開始した。12月3日に福岡国際会議場で開催された日本生物物理学会第46回年会において学会会員に対し広くこの領域研究の趣旨を伝え、そこで対象とする課題を対象としたシンポジウムを開催した。このシンポジウムには200名を超す参加者とともにこの古くて新しい問題について討論し、この問題の重要性をアピールすることができた。広報活動の一環として、本領域の活動を紹介するホームページを立ち上げた。さっそくこの生物物理学会の年会シンポジウムにおける講演概要を掲載し、本領域がめざす目標の公示と、公募研究にもとめる内容に関する情報を随時更新する態勢で提供を始めた。12月13日には東京慈恵会医科大学において開催された「筋生理の集い」において、領域研究の紹介として総括班から3 名が発表しPRを行った。1月22日には、中央大学駿河台記念館において、第1回公開シンポジウムを開催した。ここ

    researchmap

  • An experimental platform for membrane protein analysis by using micro/nano machining technologies

    Grant number:18074001  2006.4 - 2009.3

    文部科学省  科学研究費補助金(日本学術振興会・文部科学省)-特定領域研究  Grant-in-Aid for Scientific Research on Priority Areas  The University of Tokyo

    竹内 昌治

      More details

    Grant type:Competitive

    本研究では、膜超分子モータの機能解明の効率化を可能とする実験プラットフォームを、マイクロ・ナノ加工技術を利用して実現することを目的とする。具体的には、マイクロ・ナノ加工技術を利用して、膜形成のための小孔を作製し、再現性や安定性の向上を図る。膜構造の再構成技術は、各種の膜タンパク質の機能や特性の解明に必要不可欠なプラットフォームとである。また、本研究で注目する膜分子モータに限らず、イオンチャンネルや、薬剤排出とトランスポータなどへの応用も期待できる。これまでの国内外で発表されている人工平面膜法は、一分子の機能解析が可能で、膜電流が計測不能なトランスポータなどの活性計測にも有効であるが、脂質2重膜の再構成プロセスは運まかせで、再構成膜の再現性、安定性は極めて低いという問題があった。また、複数の脂質膜を同時に再構成するのは至難の業であった。 そこで、ここではマイクロ・ナノ加工技術を利用して、小型チ

    researchmap

  • 高度好塩菌の膜インターフェースにおける物質輸送の分子論的基盤

    2005.4 - 2006.3

    文部科学省  科学研究費補助金(特定領域研究) 

      More details

    Grant type:Competitive

    Grant amount: \2900000

    researchmap

  • F1モーターに外力を加えるナノシステムの構築

    2005.4 - 2006.3

    文部科学省  科学研究費補助金(特定領域研究) 

      More details

    Grant type:Competitive

    Grant amount: \2800000

    researchmap

  • バクテリオロドプシンとカーボンナノチューブの複合型マイクロナノ光センサーの構築

    2005.4 - 2006.3

    文部科学省  科学研究費補助金(萌芽研究) 

      More details

    Grant type:Competitive

    Grant amount: \1300000

    researchmap

  • Elucidation of the input-output characteristics of F_1-motor

    Grant number:14380313  2002 - 2004

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (B)  Tokyo Institute of Technology

    EIRO Muneyuki

      More details

    Grant amount: \8500000 ( Direct Cost: \8500000 )

    Biological motor proteins convert free energy of ATP hydrolysis to mechanical work, thus playing essential roles in life. But the fundamental question how they work at different free energy of ATP hydrolysis has been unanswered. This question becomes serious when we ask how stepping motion of a single motor molecule in a medium reflects the concentrations of ATP. ADP and inorganic phosphate in their individual actions. The F_1 portion of ATP synthase, also called as F_1-ATPase, is a rotary molecular motor in which central γ subunit rotates against α_3β_3 cylinder hydrolyzing ATP. Its structure is highly stable. The rotary action of this motor is processive and generates high torque. These features are ideal to explore the relationship between input free energy and output mechanical work except that this motor protein is severely inhibited by ADP. Here we overcame this problem by introducing several mutations while keeping high enzymatic activity. With a probe of beads attached, stepping rotation against viscous load was examined at a wide range of free energy by changing ADP concentration. The results showed that the work of individual step motion was not affected by the free energy of ATP hydrolysis but their frequency depended on the free energy of ATP showing a property of a molecular machine operating in an environment dominated by Brownian motion.
    In addition, we have analyzed rotations of a mutant F_1 subcomplex (E190D) which hydrolyzes ATP slowly and rotations of the wild-type F_1 employing a slowly hydrolysable substrate ATPγS (adenosine-5'-(γ-thio)-triphosphate). In both cases, interim dwells at 80° position were extended. We concluded that that cleavage of ATP takes place during the interim dwell.
    Furthermore, we could observe rotation under an optical microscope, while watching which of the three sites bound and released a fluorescent ATP analog. If we assume that the analog mimics authentic ATP, the following scheme emerges : (i)in the ATP-waiting state, one site, dictated by the orientation of γ, is empty whereas the other two bind a nucleotide ; (ii)ATP binding to the empty site drives 〜80° rotation of γ ; (iii)this triggers a reaction(s), hydrolysis and/or phosphate release but not ADP release, in the site that bound ATP one step ago ; (iv)completion of that reaction induces further 〜40° rotation.
    Thus, we could elucidate important features of the energetics and kinetics of the F1-motor.

    researchmap

  • Elucidation of the Cl-transport mechanism of halorthodopsin by time-resolved electric and spectroscopic measurements

    Grant number:11680653  1999 - 2001

    Tokyo Institute of Technology  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)  Tokyo Institute of Technology

    宗行 英朗

      More details

    Authorship:Principal investigator  Grant type:Competitive

    Grant amount: \3700000 ( Direct Cost: \3700000 )

    First of all, we improved the photovoltage measurements system so that it enabled us to examine the photovoltage generation from 1 μsec to 500 msec.
    Then, using the time-resolved photovoltage measurement system, we examined photovoltage kinetics of halorthodopsin from Halobacterium salinarum. As a function of chloride concentrations. The photovoltage consisted of two major components ; one with a sub-ms range time constant and the other with ms range time constant with different amplitudes., These components exhibited different Cl^- concentration dependencies (0.1 M to 9 M). We found that the time constant for the fast component was relatively independent of the Cl^- concentration, whereas the time constant for the slow component increased sigmoidally at higher Cl^- concentrations. The fast and the slow processes were attributed to charge (Cl^-) movements within the protein and related to Cl^- ejection, respectively. The laser photolysis studies of shR-membrane suspensions revealed that they correspond to the formation and the decay of the N intermediate. The photovoltage amplitude of the slow component exhibited a distorted bell-shaped Cl^- concentration dependence and the Cl^- concentration dependence of its time constant suggested a weak and highly cooperative Cl^- binding site(s) at cytoplasmic side (apparent K_D of about 5 M and Hill coefficient ≧ 5). The Cl^- concentration dependence of the photovoltage amplitude and the time constant for the slow process suggested a competition between spontaneous relaxation and ion translocation. The time constant for the relaxation was estimated to be > 100 ms.
    Electrogenic activity of the halorhodopsin (hR) from shark strain was also examined. After absorbing hR-overproduced membrane fraction onto a thin polymer film, we measured photoelectric current upon illumination at various Cl^- concentrations. The amplitude of the photoelectric current exhibited a bell-shaped Cl^- concentration dependency. It was found that R108Q mutant also exhibited photoelectric current at a high Cl^- concentration. The result indicates that R108 is important for uptaking Cl^- from the medium, but once Cl^- is located near the Schiff base, hR can exhibit electrogenicity even without R108.

    researchmap

  • Elucidation of the mechanism of ion transport by bacteriorhodopsin and halorhodopsin by electrical measuements

    Grant number:09833001  1997 - 1998

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research (C)  TOKYO INSTITUTE OF TECHNOLOGY

    MUNEYUKI Eiro

      More details

    Grant amount: \2700000 ( Direct Cost: \2700000 )

    In the first year, I constructed an experimental system which enabled to measure the fast photoelectric response of the membrane fractions containing bacteriorhodopsin (bR) or halorhodopsin (hR) by adsorbing them onto a thin polymer film, In the second year, I examined the photocurrent of bR and its mutant, two kinds of hRs from different halophytic bacteria as a function of pH or Cl-concentrations. The result suggested that the pH dependency of the photoelectric current by bR reflected the affinities of the entrance and exit of the proton pathway of bR for protons. similar results were obtained for hRs and it was suggested that the way of ion translocation in the pump proteins were similar to th at of a multiion channel. By using a pulsed laser flash, I could measure the charge movement within the pump protein in a single turnover. The charge movement in bR occurred in the musec and msec time region whereas the charge movement in two hRs occurred in the msec and sub-msec time region. By comparing the data obtained in the electrical measurements with the data obtained in the photochemical cycle, it was suggested that the charge movement occurs during the formation and decay of the N intermediate.
    On the other hand, I carried out a theoretical model study based on the properties of proton translocation by bR The model assumes three ion binding sites and the affinity change induce the uni-directional ion translocation. Furthermore, the distribution of the high affinity state and low affinity state must deviate from equilibrium distribution in order to induce active transport. I propose this is the critical difference between ion pumps and ion channels. The experimental results obtained in the second year fairly matches this idea. Thus it seems reasonable to assume that an ion pump is a multiion channel in which the affinity change for transported ion induces active transport.

    researchmap

  • Molecular mechanism of Energy conversion on the biomembrane

    Grant number:06045012  1994 - 1996

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for international Scientific Research  TOKYO Inst.Technology

    YOSHIDA Masasuke, MONTAL Mauricio, MUNEYUKI Eiro, ALLISON William S

      More details

    Grant amount: \5000000 ( Direct Cost: \5000000 )

    In order to know how many functional catalytic sites are necessary for ATPase activity of F1-ATPase from a thermophilic Bacillus PS3, a new method to isolate homogeneous prepartion of the alpha3beta3gamma complex with 1,2, or 3 incompetent catalytic sites was developed. Ten glutamic acids (Glu・Tag) were linked to C-terminus of the catalytically incompetent beta (E190Q) subunit. Glu・Tag itself did not affect ATPase activity of the complexes. Two kinds of alpha3beta3gamma complexes, one containing beta (wild-type) and the other Glu・Tag-linked beta (E190Q), were mixed, urea-denatured, dialyzed, and alpha3beta3gamma complexes were reconstituted. Each of the complexes containing different number of Glu・Tag-linked beta (E190Q) was separated by anion-exchange chromatography and analyzed. The results were as follows. 1) Normal steady-state ATPase activity requires three intact catalytic sites. 2) Chase-acceleration, a catalytic cooperativity, requires at least two intact catalytic sites. 3) Single-site catalysis can be mediated by a single intact catalytic site alone. Re-scrambling of subunits between complexes could occur when the complex was aged under some condition and this might be one of the reasons of the previous contradictory result (Miwa et al. (1989) J.Biochem. (Tokyo) 106,730-734).

    researchmap

  • 一分子観察によるF.モーターの回転と酵素反応ステップの関係の解明

    Tokyo Institute of Technology  Grant-in-Aid for Scientific Research on Priority Areas (A) 

    宗行 英朗

      More details

    Authorship:Principal investigator  Grant type:Competitive

    Grant amount: \2100000 ( Direct Cost: \2100000 )

    researchmap

  • 一分子観察によるF_1モーターの回転と酵素反応ステップの関係の解明

    Tokyo Institute of Technology  Grant-in-Aid for Scientific Research on Priority Areas (A) 

    宗行 英朗

      More details

    Authorship:Principal investigator  Grant type:Competitive

    Grant amount: \2100000 ( Direct Cost: \2100000 )

    researchmap

  • イオン輸送性ATP合成酵素のATPase活性とチャネル活性の共役

    Tokyo Institute of Technology  Grant-in-Aid for Scientific Research on Priority Areas 

    宗行 英朗

      More details

    Authorship:Principal investigator  Grant type:Competitive

    Grant amount: \1800000 ( Direct Cost: \1800000 )

    researchmap

▼display all

Intellectual property rights

  • 環状のナノ粒子集合体およびその製造方法

    芳賀 正明,小林 克彰,金井塚 勝彦,藤井 翔

     More details

    Application no:特願2009-213578  Date applied:2009.9.15

    Announcement no:特開2011-62607  Date announced:2011.3.31

    Applicant (Organization):学校法人中央大学

  • 鎖状分子の操作方法

    芳賀 正明,小林 克彰,金井塚 勝彦,藤井 翔

     More details

    Application no:特願2009-206143  Date applied:2009.9.7

    Announcement no:特開2011-056337  Date announced:2011.3.24

    Applicant (Organization):学校法人中央大学