Volkán-Kacsó, Sándor and Marcus, Rudolph A. (2016) Theory of single-molecule controlled rotation experiments, predictions, tests, and comparison with stalling experiments in F_1-ATPase. Proceedings of the National Academy of Sciences of the United States of America, 113 (43). pp. 12029-12034. ISSN 0027-8424. PMCID PMC5087055. https://resolver.caltech.edu/CaltechAUTHORS:20161013-110352184
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Abstract
A recently proposed chemomechanical group transfer theory of rotary biomolecular motors is applied to treat single-molecule controlled rotation experiments. In these experiments, single-molecule fluorescence is used to measure the binding and release rate constants of nucleotides by monitoring the occupancy of binding sites. It is shown how missed events of nucleotide binding and release in these experiments can be corrected using theory, with F1-ATP synthase as an example. The missed events are significant when the reverse rate is very fast. Using the theory the actual rate constants in the controlled rotation experiments and the corrections are predicted from independent data, including other single-molecule rotation and ensemble biochemical experiments. The effective torsional elastic constant is found to depend on the binding/releasing nucleotide, and it is smaller for ADP than for ATP. There is a good agreement, with no adjustable parameters, between the theoretical and experimental results of controlled rotation experiments and stalling experiments, for the range of angles where the data overlap. This agreement is perhaps all the more surprising because it occurs even though the binding and release of fluorescent nucleotides is monitored at single-site occupancy concentrations, whereas the stalling and free rotation experiments have multiple-site occupancy.
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Alternate Title: | Theory of single-molecule controlled rotation experiments, predictions, tests, and comparison with stalling experiments in F1-ATPase | |||||||||||||||
Additional Information: | © 2016 National Academy of Sciences. Contributed by Rudolph A. Marcus, August 26, 2016 (sent for review July 15, 2016; reviewed by Attila Szabo and Arieh Warshel). Published online before print October 10, 2016, doi: 10.1073/pnas.1611601113 We thank Drs. Imre Derényi and Kengo Adachi for helpful discussions and comments and the reviewers for useful suggestions. This work was supported by the Office of the Naval Research, the Army Research Office, and the James W. Glanville Foundation. Author contributions: S.V.-K. and R.A.M. designed research, performed research, analyzed data, and wrote the paper. Reviewers: A.S., National Institutes of Health; and A.W., University of Southern California. The authors declare no conflict of interest. This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1611601113/-/DCSupplemental. | |||||||||||||||
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Subject Keywords: | F1-ATPase; biomolecular motors; single-molecule imaging; nucleotide binding; group transfer theory | |||||||||||||||
Issue or Number: | 43 | |||||||||||||||
PubMed Central ID: | PMC5087055 | |||||||||||||||
Record Number: | CaltechAUTHORS:20161013-110352184 | |||||||||||||||
Persistent URL: | https://resolver.caltech.edu/CaltechAUTHORS:20161013-110352184 | |||||||||||||||
Official Citation: | Sándor Volkán-Kacsó and Rudolph A. Marcus Theory of single-molecule controlled rotation experiments, predictions, tests, and comparison with stalling experiments in F1-ATPase PNAS 2016 113 (43) 12029-12034; published ahead of print October 10, 2016, doi:10.1073/pnas.1611601113 | |||||||||||||||
Usage Policy: | No commercial reproduction, distribution, display or performance rights in this work are provided. | |||||||||||||||
ID Code: | 71049 | |||||||||||||||
Collection: | CaltechAUTHORS | |||||||||||||||
Deposited By: | George Porter | |||||||||||||||
Deposited On: | 14 Oct 2016 21:12 | |||||||||||||||
Last Modified: | 22 Nov 2019 09:58 |
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