DOI: 10.1073/pnas.95.14.8034. Myosin conformational states determined by single fluorophore polarization

Myosin conformational states determined by single fluorophore polarization

10.1073/pnas.95.14.8034

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

Muscle contraction is powered by the interaction of the molecular motor myosin with actin. With new techniques for single molecule manipulation and fluorescence detection, it is now possible to correlate, within the same molecule and in real time, conformational states and mechanical function of myosin. A spot-confocal microscope, capable of detecting single fluorophore polarization, was developed to measure orientational states in the smooth muscle myosin light chain domain during the process of motion generation. Fluorescently labeled turkey gizzard smooth muscle myosin was prepared by removal of endogenous regulatory light chain and re-addition of the light chain labeled at cysteine-108 with the 6-isomer of iodoacetamidotetramethylrhodamine (6-IATR). Single myosin molecule fluorescence polarization data, obtained in a motility assay, provide direct evidence that the myosin light chain domain adopts at least two orientational states during the cyclic interaction of myosin with actin, a randomly disordered state, most likely associated with myosin whereas weakly bound to actin, and an ordered state in which the light chain domain adopts a finite angular orientation whereas strongly bound after the powerstroke.

Bibliography

Warshaw, D. M., Hayes, E., Gaffney, D., Lauzon, A.-M., Wu, J., Kennedy, G., Trybus, K., Lowey, S., & Berger, C. (1998). Myosin conformational states determined by single fluorophore polarization. Proceedings of the National Academy of Sciences, 95(14), 8034â8039.

Authors 9

David M. Warshaw (first)
Eric Hayes (additional)
Donald Gaffney (additional)
Anne-Marie Lauzon (additional)
Junru Wu (additional)
Guy Kennedy (additional)
Kathleen Trybus (additional)
Susan Lowey (additional)
Christopher Berger (additional)

References 30 Referenced 102

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Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 10:39 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 4:40 p.m.)
Indexed	1 month, 3 weeks ago (July 8, 2025, 7:29 a.m.)
Issued	27 years, 1 month ago (July 7, 1998)
Published	27 years, 1 month ago (July 7, 1998)
Published Online	27 years, 1 month ago (July 7, 1998)
Published Print	27 years, 1 month ago (July 7, 1998)

Funders 0

None

BibTeX

@article{Warshaw_1998, title={Myosin conformational states determined by single fluorophore polarization}, volume={95}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.95.14.8034}, DOI={10.1073/pnas.95.14.8034}, number={14}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Warshaw, David M. and Hayes, Eric and Gaffney, Donald and Lauzon, Anne-Marie and Wu, Junru and Kennedy, Guy and Trybus, Kathleen and Lowey, Susan and Berger, Christopher}, year={1998}, month=jul, pages={8034–8039} }

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