Abstract
The measurement of polarized fluorescence from a fluorescent ATP or ADP analog, ɛ-ATP or ɛ-ADP (1, N8 -Etheno-ATP or ADP) bound to myosin heads in a glycerinated muscle fiber revealed that during isometric contraction at low concentrations of Mg-ɛ-ATP the bound nucleotides are highly oriented with respect to the fiber axis, and their mean angle is almost the same as that in rigor. Furthermore, the polarization of tryptophan fluorescence did not change when the fiber was transferred from a rigor state to an active state at low ATP concentrations. Thus, if a rotation of myosin heads occurs during contraction, it seems to be very limited. The angle of bound ɛ-ADP is not changed by passive stretching of the fiber in the presence of ɛ-ADP.
By using the same technique, it was found that orientation of a phalloidin-FITC complex (Ph-FTTC) specifically bound to F-actin in a glycerinated muscle fiber is significantly changed when the fiber is activated from relaxation or rigor to contraction. No change in orientation of Ph-FITC bound to F-actin is induced by simple addition of Mg-ADP or by passive stretching of the fiber in the absence of ATP. These results suggest that a rotation or a distortion of actin monomers occurs during contraction, which is involved in the process of active tension development.
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References
Balint, M., Wolf, I., Tarcsafavi, A., Gergely, J.P., Sreter, A. (1978). Location of SH-1 and SH-2 in the heavy chain segment of meromyosin. Arch. Biochem. Biophys. 190: 793–799.
Cooke, R. (1981). Stress does not alter the conformation of a domain of the myosin cross-bridge in rigor muscle fibers. Nature 294: 570–571.
Cooke, R., Crowder, M., and Thomas, D.D. (1982). Measuring cross-bridge angles with paramagnetic probes in rigor, relaxed and contracting muscle fibers. In: Cross-bridge Mechanisms in Muscle Contraction. eds. G.H. Pollack and H. Sugi.
Denker, P., Low, J., Hassellach, W. and Wieland, T. (1975). Interaction of actin with phalloidin: Polymerization and staloilization of F-actin. Biochim. Biophys. Acta 400: 407–414.
Huxley, A.F. (1974). Review lecture Muscle Contraction. J. Physiol. 243: 1–43.
Huxley, A.F. and Simmons, R.M. (1971). Proposed mechanism of force generation in striated muscle. Nature 233: 533–538.
Huxley, H.E. (1969). The mechanism of muscular contraction: Recent stuctural studies suggest a revealing model for cross-bridge action at variable filament spacing. Science 164: 1356–1366.
Lengsfeld, A.M., Low, J., Wieland, T., Dancker, P. and Hassellbach, W. (1974). Interaction of phalloidin with actin. Proc. Natl. Acad. Sci. USA 71: 2803–2807.
Mornet, D., Bertrand, R., Pantel, P., Audemard, E. and Kassab, R. (1981). Structure of the actin-myosin interface. Nature 292: 301–306.
Onishi, H., Ohtsuka, E., Ikehara, M. and Tonomura, Y. (1973). Energy transfer from tryptophan residues to a fluorescent ATP analog, 1, N8-Ethenoadenosine triphosphate, bound to H-meromyosin. J. Bloch. 74: 435–450.
Oosawa, F. (1977). Actin-actin bond strength and the conformational change of F-actin. Biorheology 14: 11–19.
Oosawa, F. (1980). Dynamics of Actin Filament. In: Muscle Contraction: Its Regulatory Mechanisms. Ebashi, S, Maruyama, K and Endo, M., eds. Japan Science Societies Press/Springer-Verlag. pp. 165–172.
Sekine, T. and Yamaguchi, M. (1963). Effect of ATP on the binding of N-ethylmaleimide to SH groups in the active site of myosin ATPase. J. Biochem. 54: 196–198.
Szilagyi, L., Balint, M., Sneter, F.A. and Gergely, J. (1979). Photo affinity labelling with an ATP analog of the N-terminal peptide of myosin. Biochem. Biophys. Res. Comm. 87: 936–945.
Thomas, D.D. and Cooke, R. (1980). Orientation of spin-labeled myosin heads in glycerinated muscle fibers. Biophys. J. 32: 891–906.
Thomas, D.D., Ishíwata, S.I., Seidel, J.C. and Gergely, J. (1980) Rotational dynamics of spin-labeled myosin heads in myofibrils. Biophys. J. 32: 873–890.
Tonomura, Y. (1972). Muscle Proteins, Muscle Contraction and Cation Transport. Tokyo: University of Tokyo Press.
Wulf, E., Deboben, A., Bautz, F.A., Faulstich, H. and Wieland, T. (1979). Fluorescent phallotoxin, a tool for the visualization of cellular actin. Proc. Natl. Acad. Sci. USA 76: 4498–4502.
Yanagida, T. (1980). The angles of cross-bridges bound to nucleotide at various concentrations of Mg-(ɛ)-ATP In: Muscle Contraction: Its Regulatory Mechanisms Ebashi et al., eds. pp. 165–172.
Yanagida, T. (1981a). Angles of nucleotides bound to cross-bridges in glycerinated muscle fiber at various concentrations of ɛ-ATP, ɛ-ADP and ɛ-AMPPNP detected by polarized fluorescence. J. Mol. Biol. 148: 539–560.
Yanagida, T. (1981b). Rotational motion of actin monomers in a muscle fiber during contraction. VH International Biophys. Cong. Abs. p. 274.
Yanagida, T. and Oosawa, F. (1978). Polarized fluorescence from E-ADP incorporated into F-actin in a myosin-free single fiber: Conformation of F-actin and changes induced in it by heavy meromyosin. J. Mol. Biol. 126: 507–524.
Yanagida, T. and Oosawa, F. (1980). Conformtional changes in their filaments in muscle fibers induced by Caz2 + ions. J. Mol. Biol. 140: 313–320.
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© 1984 Plenum Press, New York
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Yanagida, T. (1984). Angles of Fluorescently Labelled Myosin Heads and Actin Monomers in Contracting and Rigor Stained Muscle Fiber. In: Pollack, G.H., Sugi, H. (eds) Contractile Mechanisms in Muscle. Advances in Experimental Medicine and Biology, vol 37. Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-4703-3_36
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DOI: https://doi.org/10.1007/978-1-4684-4703-3_36
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