DOI: 10.1242/jeb.203.1.1. Rotation of F1-ATPase and the hinge residues of the β subunit

Rotation of F1-ATPase and the hinge residues of the β subunit

10.1242/jeb.203.1.1

Crossref journal-article

The Company of Biologists

Journal of Experimental Biology (237)

Abstract

ABSTRACT Rotation of a motor protein, F1 -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 β subunit. We have visualised rotation of a single molecule of F1 -ATPase by attaching a fluorescent actin filament to the top of the γ subunit in the immobilised F1 -ATPase, thus settling a major controversy regarding the rotary catalysis. The rotation of the γ subunit was exclusively in one direction, as could be predicted by the crystal structure of bovine heart F1 -ATPase. Rotation at low ATP concentrations revealed that one revolution consists of three 120 ° steps, each fuelled by the binding of an ATP to the β subunit. The mean work done by a 120 ° 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 β subunit, which undergoes large opening–closing domain motion upon binding of AT(D)P. We identified three hinge residues, βHis179, βGly180 and βGly181, 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 β 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.

Bibliography

Masaike, T., Mitome, N., Noji, H., Muneyuki, E., Yasuda, R., Kinosita, K., & Yoshida, M. (2000). Rotation of F1-ATPase and the hinge residues of the Î² subunit. Journal of Experimental Biology, 203(1), 1â8.

Authors 7

Tomoko Masaike (first)
Noriyo Mitome (additional)
Hiroyuki Noji (additional)
Eiro Muneyuki (additional)
Ryohei Yasuda (additional)
Kazuhiko Kinosita (additional)
Masasuke Yoshida (additional)

References 22 Referenced 50

10.1038/370621a0 / Nature / Structure at 2.8 Å resolution of F1-ATPase from bovine heart mitochondria by Abrahams (1994)
10.1074/jbc.273.2.865 / J. Biol. Chem / ATP synthesis by FoF1-ATP synthase independent of noncatalytic nucleotide binding sites and insensitive to azide inhibition by Bald (1998)
10.1016/0005-2728(93)90063-L / Biochim. Biophys. Acta / The binding change mechanism for ATP synthase – some probabilities and possibilities by Boyer (1993)
{'key': '2024121803531574100_JEXBIO_203_1_1C4', 'first-page': '231', 'article-title': 'The present status of the binding-change mechanism and its relation to ATP formation by chloroplasts', 'volume-title': 'In Energy Coupling in Photosynthesis', 'author': 'Boyer', 'year': '1981'} / In Energy Coupling in Photosynthesis / The present status of the binding-change mechanism and its relation to ATP formation by chloroplasts by Boyer (1981)
10.1073/pnas.92.24.10964 / Proc. Natl. Acad. Sci. USA / Rotation of subunits during catalysis by Escherichia coli F1-ATPase by Duncan (1995)
10.1021/bi00028a004 / Biochemistry / X-ray structures of the myosin motor domain of Dictyostelium discoideum complexed with MgADP·BeFx and MgADP·AlF4− by Fisher (1995)
{'key': '2024121803531574100_JEXBIO_203_1_1C7', 'first-page': '19', 'article-title': 'Structural studies of myosin:nucleotide complexes: a revised model for the molecular basis of muscle contraction', 'volume': '68', 'author': 'Fisher', 'year': '1995', 'journal-title': 'Biophys. J'} / Biophys. J / Structural studies of myosin:nucleotide complexes: a revised model for the molecular basis of muscle contraction by Fisher (1995)
10.1006/bbrc.1994.1336 / Biochim. Biophys. Res. Commun / Single-molecule analysis of the actomyosin motor using nano-manipulation by Ishijima (1995)
10.1021/bi00050a023 / Biochemistry / The α3β3γ complex of the F1-ATPase from the thermophilic Bacillus PS3 containing the α-D261N substitution fails to dissociate inhibitory MgADP from a catalytic site when ATP binds to noncatalytic sites by Jault (1995)
10.1074/jbc.273.31.19375 / J. Biol. Chem / Direct observation of the rotation of ε subunit in F1-ATPase by Kato-Yamada (1998)
10.1038/379311a0 / Nature / The 2.0 Å crystal structure of a heterotrimeric G protein by Lambright (1996)
10.1016/S0021-9258(18)48416-5 / J. Biol. Chem / The G226A mutant of Gsα highlights the requirement for dissociation of G protein subunits by Lee (1992)
10.1074/jbc.272.13.8215 / J. Biol. Chem / Catalytic activity of the α3β3γ complex of F1-ATPase without noncatalytic nucleotide binding site by Matsui (1997)
10.1038/366654a0 / Nature / The 2.2 Å crystal structure of transducin-α complexed with GTPγS by Noel (1993)
10.1038/386299a0 / Nature / Direct observation of the rotation of F1-ATPase by Noji (1997)
10.1038/381623a0 / Nature / Intersubunit rotation in active F1-ATPase by Sabbert (1996)
10.1074/jbc.273.32.20334 / J. Biol. Chem / Mutational analysis of the switch II loop of Dictyostelium myosin II by Sasaki (1998)
10.1021/bi952633+ / Biochemistry / X-ray structure of the magnesium(II). ADP. vanadate complex of the Dictyostelium discoideum myosin motor domain to 1.9 Å resolution by Smith (1996)
10.1038/365721a0 / Nature / Direct observation of kinesin stepping by optical trapping interferometry by Svoboda (1993)
10.1016/S0005-2728(98)00150-9 / Biochim. Biophys. Acta / Contraction transition of F1-Fo ATPase during catalytic turnover by Syroeshkin (1998)
10.1016/0014-5793(95)01276-2 / FEBS Lett / Conformational dynamics monitored by His-179 and His-200 of isolated thermophilic F1-ATPase β subunit which reside at the entrance of the conical tunnel in holoenzyme by Tozawa (1995)
10.1016/S0092-8674(00)81456-7 / Cell / F1-ATPase is a highly efficient molecular motor that rotates with discrete 120 ° steps by Yasuda (1998)

Dates

Type	When
Created	4 years, 4 months ago (April 25, 2021, 1:33 a.m.)
Deposited	8 months, 1 week ago (Dec. 17, 2024, 10:53 p.m.)
Indexed	1 month, 3 weeks ago (July 1, 2025, 11:48 a.m.)
Issued	25 years, 7 months ago (Jan. 1, 2000)
Published	25 years, 7 months ago (Jan. 1, 2000)
Published Online	25 years, 7 months ago (Jan. 1, 2000)
Published Print	25 years, 7 months ago (Jan. 1, 2000)

Funders 0

None

BibTeX

@article{Masaike_2000, title={Rotation of F1-ATPase and the hinge residues of the β subunit}, volume={203}, ISSN={1477-9145}, url={http://dx.doi.org/10.1242/jeb.203.1.1}, DOI={10.1242/jeb.203.1.1}, number={1}, journal={Journal of Experimental Biology}, publisher={The Company of Biologists}, author={Masaike, Tomoko and Mitome, Noriyo and Noji, Hiroyuki and Muneyuki, Eiro and Yasuda, Ryohei and Kinosita, Kazuhiko and Yoshida, Masasuke}, year={2000}, month=jan, pages={1–8} }

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  "abstract": "<jats:title>ABSTRACT</jats:title>\n               <jats:p>Rotation of a motor protein, F1 -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 \u03b2 subunit. We have visualised rotation of a single molecule of F1 -ATPase by attaching a fluorescent actin filament to the top of the \u03b3 subunit in the immobilised F1 -ATPase, thus settling a major controversy regarding the rotary catalysis. The rotation of the \u03b3 subunit was exclusively in one direction, as could be predicted by the crystal structure of bovine heart F1 -ATPase. Rotation at low ATP concentrations revealed that one revolution consists of three 120 \u00b0 steps, each fuelled by the binding of an ATP to the \u03b2 subunit. The mean work done by a 120 \u00b0 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 \u03b2 subunit, which undergoes large opening\u2013closing domain motion upon binding of AT(D)P. We identified three hinge residues, \u03b2His179, \u03b2Gly180 and \u03b2Gly181, 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 \u03b2 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.</jats:p>",
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    "date-parts": [
      [
        2000,
        1,
        1
      ]
    ]
  }
}