Abstract
Based on techniques for single molecule imaging and nanomanipulation by optical tweezers, we have developed a new technique that allows simultaneous measurement of individual ATPase and mechanical reactions from a single myosin molecule during force generation. We show how the ATPase reaction couples to the mechanical reaction directly at the single molecule level. The results show that the myosin head can produce force even after releasing the bound nucleotide, probably ADP, suggesting that the chemical energy driven by ATP hydrolysis can be hysteretically stored in the myosin molecule. This view does not support a widely accepted hypothesis in which the force generation is tightly coupled to ligand dissociation.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
T. Yanagida, M. Nakase, K. Nishiyama and F. Oosawa: Nature (London) 307 (1984) 58.
S.J. Kron and J.A. Spudich: Proc. Natl. Acad. Sci. USA 83 (1986) 6272.
Y. Harada, A. Noguchi, A. Kishino and T. Yanagida: Nature (London) 326 (1987) 805.
H.E. Huxley: J. Biol. Chem. 265 (1990) 8347.
A. Kishino and T. Yanagida: Nature (London) 334 (1988) 74.
A. Ishijima, T. Doi, K. Sakurada and T. Yanagida: Nature (London) 352 (1991) 301.
A. Ishijima, H. Kojima, H. Higuchi, Y. Harada, T. Funatsu and T. Yanagida: Biophys. J. 70 (1996) 383.
A. Ishijima, Y. Harada, H. Kojima, T. Funatsu, H. Higuchi and T. Yanagida: Biochem. Biophys. Res. Commun. 199 (1994) 1057.
J.T. Finer, R.M. Simmons and J.A. Spudich: Nature (London) 368 (1994) 113.
R.W. Lymn and E.W. Taylor: Biochemistry 10 (1971) 4617.
T. Funatsu, Y. Harada, M. Tokunaga, K. Saito and T. Yanagida: Nature (London) 374 (1995) 555.
R.D. Vale, T. Funatsu, D.W. Pierce, L. Romberg, Y. Harada and T. Yanagida: Nature (London) 380 (1996) 451.
A. Ishijima, H. Kojima, T. Funatsu, M. Tokunaga, H. Higuchi, H. Tanaka and T. Yanagida: Cell 92 (1998) 161.
J.E. Molloy, J.E. Burns, J. Kendrick-Jones, R.T. Tregear and D.C.S. White: Nature (London) 378 (1995) 209.
A.F. Huxley and R.M. Niedergerke: Nature (London) 173 (1954) 971.
H.E. Huxley and J. Hanson: Nature (London) 173 (1954) 973.
H.E. Huxley: Science 164 (1969) 1356.
A.F. Huxley and R.M. Simmons: Nature (London) 233 (1971) 533.
T. Kodama: Physiol. Rev. 65 (1985) 467.
T. Ha, T. Enderle, D.F. Ogletree, D.S. Chemla, P.R. Selvin and S. Weiss: Proc. Natl. Acad. Sci. USA 93 (1996) 6264.
R.M. Dickson, A.B. Cubitt, R.Y. Tsien and W.E. Moerner: Nature (London) 24 (1997) 355.
C. Frieden: J. Biol. Chem. 245 (1970) 5788.
Y. Harada, K. Sakurada, T. Aoki, D.D. Thomas and T. Yanagida: J. Mol. Biol. 216 (1990) 49.
M. Tokunaga, K. Kitamura, K. Saito, A. H. Iwane and T. Yanagida: Biochem. Biophys. Res. Commun. 235 (1997) 47.
K. Svoboda, C.F. Schmidt, B.J. Schnapp and S.M. Block: Nature (London) 365 (1993) 721.
Author information
Authors and Affiliations
Corresponding author
Additional information
This paper was originally presented as an invited paper at the seventh Meeting on Near Field Optics, which was held on July 1, 1998 at Nagoya University, Nagoya, organized by the Research Group on Near Field Optics of the Optical Society of Japan, an affiliate of the Japan Society of Applied Physics.
Rights and permissions
About this article
Cite this article
Ishijima, A., Kojima, H., Tanaka, H. et al. Simultaneous Measurement of Individual ATPase and Mechanical Reactions by a Single Myosin Molecule at Work. OPT REV 6, 16–23 (1999). https://doi.org/10.1007/s10043-999-0016-5
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s10043-999-0016-5