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Changes in anisotropy of tryptophan fluorescence and in birefringence of actin filaments induced by the binding to actin of heavy meromyosin (HMM), both containing DTNB light chains and devoid of them, were found in rabbit muscle fibres free of myosin, troponin, and tropomyosin. Ca2+ was shown to affect the pattern of changes in tryptophan fluorescence anisotropy and birefringence of actin filament at the moment of HMM interaction with actin, providing HMM contains DTNB light chains. Anisotropy of tryptophan fluorescence and birefringence of actin filaments rises in the absence of Ca2+ (pCa greater than or equal to 7), while in its presence (pCa less than or equal to 6) these values drop down. Furthermore, these changes become cooperative when Ca2+ concentration increases from pCa = 7 to pCa = 6. It was shown that the binding of HMM devoid of DTNB light chains to F-actin decreases tryptophan fluorescence anisotropy and birefrigence of actin filaments, regardless of Ca2+ concentration. Ca2+-dependent structural changes of F-actin induced by interaction of heads of myosin molecules with actin are assumed to be of great importance in regulation of muscle contraction of vertebrate skeletal muscles.

The dependence of F-actin conformational changes induced by the F-actin-HMM complex on pH and ionic strength was found by polarized ultraviolet fluorescence microscopy. It is discovered that pH affects sufficiently the cooperativity of F-actin structural changes, while the ionic strength affects their depth. The actomyosin complex was supposed to be at least in two structural states, differing in their orientation as well as in flexibility of F-actin monomers.

The polarized fluorescence of intrinsic tryptophan residues and the birefringence of ghost muscle fibres of rabbit were measured during thin filaments binding to heavy meromyosin containing 5,5'-dithiobis [2-nitrobenzoic acid] light chains and to those devoid of them with a view of investigating conformational changes in F-actin. Ca2+ binding to heavy meromyosin containing 5,5'-dithiobis [2-nitrobenzoic acid] light chains was shown to affect the character of these changes during the formation of the F-actin - heavy meromyosin complex.

Isotope-exchange reactions (H2(18)O in equilibrium with KH2PO4) during ATP hydrolysis catalyzed by myosin and its subfragment 1 from rabbit, dog, and human cardiac muscle were studied. All preparations of myosin and subfragment 1 in the presence of Mg2+ catalyzed two types of 18O-exchange reactions similar to those of skeletal muscle: intermediate and direct 18O exchange. The dependences of both reactions on divalent metals and nucleotides were studied. Data on 18O exchange of subfragment 1 from rabbit, dog, and human cardiac muscle were obtained for the first time. They indicate the similarity of molecular mechanisms of ATP energy use in cardiac and skeletal muscle contraction.

18O-exchange reactions of skeletal, cardiac and smooth muscle myosin were studied. All investigated preparations of myosin catalyse two types of 18O-exchange--intermediate and direct exchanges in the presence of Mg2+. The dependence of 18O-exchange extent on divalent cations have appeared to be similar for different muscle types. This supports the hypothesis of similarity of ATP hydrolysis molecular mechanisms by myosin of different origin.

Myosin, HMM and HMM S1 catalyze 18O-exchange between P1 and H218O of the medium at an intermediate stage of ATP hydrolysis ("intermediate 18O-exchange") in the presence of Mg2+. Natural complexes of actomyosin and acto-HMM S1 do not catalyze intermediate 18O-exchange but facilitate "direct" or "medium" 18O-exchange (KH2P18O4 in equilibrium H2O) even without ATP. Reconstituted complexes of actomyosin, acto-HMM, acto-HMM S1, PABC-HMM S1, congo-myosin and TNP-myosin do not catalyze direct 18O-exchange in the presence of Mg2+ and absence of ATP. From the data obtained a hypothetical sequence of phosphorylation and 18O-exchange reactions in myofibril action has been suggested.