Calcium binding to rabbit skeletal myosin under physiological conditions.

At a free Mg2+ concentration of 1.0 mM, myosin binds one Ca2+ per molecule when the Ca2+ concentration is 20 muM, a value in the concentration range expected during contraction of skeletal muscle. Mg2+ alters Ca2+ binding in a complex manner, not by simple competition. In the range from 20 to 100 muM Mg2+ it produces positive cooperativity between the high-affinity Ca2+ binding sites, in addition to shifting binding to higher Ca2+ concentrations. High-affinity Ca2+ binding is not significantly affected by the addition of ATP, increase in ionic strength to 0.1 and changes in temperature. Ca2+ binding did not increase actin-activated ATPase activity in the absence of regulatory proteins, but rather inhibited it.