Abstract
In an attempt to identify the binding sites of myosin fragments to the nitrocellulose film in a flow cell, proteolytic digestion was performed of myosin fragments bound to the substrate. Chymotryptic digestion of papain•Mg-subfragment-1 (S1) showed that a large proportion of S1 were bound near the tail and that the digestion site was protected for some S1. The binding of S1 near the tail was also confirmed by the production of extra long subfragment-2 (S2) by chymotryptic digestion of heavy meromyosin (HMM). Papain digestion of HMM released slightly smaller HMM from the substrate and impaired the motility at the same time. Hence, the motility of HMM is supported largely by those bound to the substrate near the C terminus. These results demonstrate the importance of flexible regions of myosin in generating active movements.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Toyoshima, Y.Y. et al. Nature 328, 536–539 (1987).
Kron, S.J., Toyoshima, Y.Y., Uyeda, T.Q.P. & Spudich, J.A. Methods Enzymol. 196, 399–416 (1990).
Manstein, DJ., Ruppel, K.M. & Spudich, J.A. Science 246, 656–658 (1989).
Collins, K., Sellers, J.R. & Matsudaira, P. J. CellBiol. 110, 1137–1147 (1990).
Schwyter, D.H., Kron, S.J., Toyoshima, Y.Y., Spudich, J.A. & Reisler, E. J. CellBiol. 111, 465–470 (1990).
Sutoh, K., Ando, M. Sutoh, K. & Toyoshima, Y.Y. Proc. Natl. Acad. Sci. USA 88, 7711–7714 (1991).
Huxley, H.E. Science 164, 1356–1366 (1969).
Toyoshima, Y.Y., Kron, S.J. & Spudich, J.A. Proc. Natl Acad. Sci. USA 87, 7130–7134 (1990).
Harada, Y., Sakurada, K., Aoki, T., Thomas, D.D. & Yanagida, T. J. Mol. Biol. 216, 49–68 (1990).
Uyeda, T.Q.P., Krön, S.J. & Spudich, J.A. J. Mol. Biol. 214, 699–710 (1990).
Ishijima, A., Doi, T., Sakurada, K. & Yanagida, T. Nature 352, 301–306 (1991).
Uyeda, T.Q.P., Warrick, H.M., Krön, S.J. & Spudich, J.A. Nature 352, 307–311 (1991).
Toyoshima, Y.Y., Toyoshima, C. & Spudich, J.A. Nature 341, 154–156 (1989).
Toyoshima, Y.Y. J. Cell Sci. suppl. 14, 83–85 (1991).
Ueno, H. & Harrington, W.F. J. Mol. Biol. 180, 667–701 (1984).
Kinosita, K., Ishiwata, S., Yoshimura, H., Asai, H. & Ikegami, A. Biochemistry 23, 5963–5975 (1984).
Miyanishi, T., Toyoshima, C., Wakabayashi, T. & Matsuda, G. J. Biochem. (Tokyo) 103, 458–462 (1988).
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1993 Springer Science+Business Media New York
About this chapter
Cite this chapter
Toyoshima, Y.Y. (1993). How are Myosin Fragments Bound to Nitrocellulose Film ?. In: Sugi, H., Pollack, G.H. (eds) Mechanism of Myofilament Sliding in Muscle Contraction. Advances in Experimental Medicine and Biology, vol 332. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-2872-2_25
Download citation
DOI: https://doi.org/10.1007/978-1-4615-2872-2_25
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4613-6245-6
Online ISBN: 978-1-4615-2872-2
eBook Packages: Springer Book Archive