Imaging of single fluorescent molecules and individual atp turnovers by single myosin molecules in aqueous solution

ABSTRACT VISUALIZATION of single actin filaments by fluorescence microscopy1led to the development of new _in vitro_ assays for analysing actomyosin-based motility at the molecular level2-5.

The ability to manipulate actin filaments with a microneedle6,7 or an optical trap8 combined with position-sensitive detectors has enabled direct measurements of nanometre displacements and

piconewton forces exerted by individual myosin molecules. To elucidate how myosin generates movement, it is necessary to understand how ATP hydrolysis is coupled to mechanical work at the

level of the single molecule. But the most sensitive microscopic ATPase assay available still requires over 1,000 myosins9. To enhance the sensitivity of such assays, we have refined

epifluorescence and total internal reflection microscopies to visualize single fluorescent dye molecules. We report here that this approach can be used directly to image single fluorescently

labelled myosin molecules and detect individual ATP turnover reactions. In contrast to previously reported single fluorescent molecule imaging methods, which used specimens immobilized on

an air-dried surface10-12, our method allows video-rate imaging of single molecules in aqueous solution, and hence can be applied to the study of many types of enzymes and biomolecules.

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support SIMILAR CONTENT BEING VIEWED BY OTHERS SINGLE MOLECULE TURNOVER OF FLUORESCENT ATP BY MYOSIN AND ACTOMYOSIN UNVEIL ELUSIVE ENZYMATIC MECHANISMS Article Open access 13 January 2021

FAST SINGLE-PARTICLE TRACKING OF MEMBRANE PROTEINS COMBINED WITH SUPER-RESOLUTION IMAGING OF ACTIN NANODOMAINS Article Open access 03 April 2025 A FUNCTIONAL FAMILY OF FLUORESCENT NUCLEOTIDE

ANALOGUES TO INVESTIGATE ACTIN DYNAMICS AND ENERGETICS Article Open access 22 January 2021 REFERENCES * Yanagida, T., Nakase, M., Nishiyama, K. & Oosawa, F. _Nature_ 307, 58–60 (1984).

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AFFILIATIONS * BioMotron Project, ERATO, JRDC, Senba-higashi 2-4-14, Mino, Osaka, 562, Japan Takashi Funatsu, Yoshie Harada, Makio Tokunaga, Kiwamu Saito & Toshio Yanagida * Department

of Biophysical Engineering, Osaka University, Toyonaka, Osaka, 560, Japan Toshio Yanagida Authors * Takashi Funatsu View author publications You can also search for this author inPubMed 

Google Scholar * Yoshie Harada View author publications You can also search for this author inPubMed Google Scholar * Makio Tokunaga View author publications You can also search for this

author inPubMed Google Scholar * Kiwamu Saito View author publications You can also search for this author inPubMed Google Scholar * Toshio Yanagida View author publications You can also

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single fluorescent molecules and individual ATP turnovers by single myosin molecules in aqueous solution. _Nature_ 374, 555–559 (1995). https://doi.org/10.1038/374555a0 Download citation *

Received: 19 December 1994 * Accepted: 08 February 1995 * Issue Date: 06 April 1995 * DOI: https://doi.org/10.1038/374555a0 SHARE THIS ARTICLE Anyone you share the following link with will

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