(32P) phosphate incorporation into ATP during ATP hydrolysis and its dependence on the interaction of actin and myosin.

The incorporation of 32Pi into ATP has been found to be catalyzed by myosin only when and if it interacts with actin. This exchange reaction is inhibited in natural but not in desensitized actomyosin after removing of trace Ca2+ with ethyleneglycol bis(2-aminoethyl)-N,N'-tetraacetic acid (EGTA). In desensitized as well as in synthetic actomyosin the exchange reaction can be fully inhibited by the addition of troponin I (0.5 mg troponin I/mg actomyosin results in a 50% inhibition) or after replacing the Mg activator by CaCl2. The exchange rate is about 1:500 of the ATPase rate in presence of 2 mM phosphate. These results suggest the existence of an 'energy-rich' actin -- myosin -- nucleoside-diphosphate intermediate during the cross-bridge cycle.