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

Shimabukuro, K., Yasuda, R., Muneyuki, E., Hara, K. Y., Kinosita, K., Yoshida, M. (December 2003) Catalysis and rotation of F-1 motor: Cleavage of ATP at the catalytic site occurs in 1 ms before 40 degrees substep rotation. Proceedings of the National Academy of Sciences of the United States of America, 100 (25). pp. 14731-14736. ISSN 0027-8424

URL: http://www.ncbi.nlm.nih.gov/pubmed/14657340
DOI: 10.1073/pnas.2434983100

Abstract

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.

Item Type: Paper
Uncontrolled Keywords: APPARENT NEGATIVE COOPERATIVITY HEART MITOCHONDRIAL ATPASE C-SUBUNIT OLIGOMER ESCHERICHIA-COLI BETA-SUBUNIT THERMOPHILIC BACTERIUM GLUTAMIC-ACID GAMMA-SUBUNIT WILD-TYPE F-1-ATPASE
Subjects: bioinformatics > genomics and proteomics > small molecules > ATP
bioinformatics > genomics and proteomics > genetics & nucleic acid processing
bioinformatics > genomics and proteomics
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types
bioinformatics > genomics and proteomics > small molecules
CSHL Authors:
Communities: CSHL labs > Svoboda lab
Depositing User: Matt Covey
Date: December 2003
Date Deposited: 01 Apr 2013 19:00
Last Modified: 01 Apr 2013 19:00
PMCID: PMC299784
Related URLs:
URI: http://repository.cshl.edu/id/eprint/28013

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