ELECTROSTATIC CONTRIBUTIONS TO THE BINDING OF MYOSIN AND MYOSIN-MGADP TO F-ACTIN IN SOLUTION

The ionic strength dependence of skeletal myosin subfragment 1 (S1) binding to unregulated F-actin was measured in solutions containing from 0 to 0.50 M added lithium acetate (LiOAc) in the absence and presence of MgADP. The data were analyzed by using a theory based on an ion interaction model that is rigorous for high ionic strength solutions [Pitzer, K. S. (1973) J. Phys. Chem, 77, 268-277] in order to obtain values for K, the equilibrium association constant when the ionic strength is zero, and for |zMzA|, the absolute value of the product of the net electric charges of the actin binding site on myosin (zM) and the myosin binding site on actin (zA). The presence of MgADP reduced K by a factor of 10, as expected, and reduced |zMzA| by about 1 esu2. Because the presence of MgADP is not likely to change the net charge of the myosin binding site on actin, these data are consistent with a model in which MgADP binding to S1 reduces its affinity for actin by a mechanism that reduces the net electric charge of the acting binding site on S1. The value of |zMzA| in the absence of ADP was 8.1 ± 0.9 esu2, which, if one uses integer values, suggests that zM and zA are in the 8+ to 1+ esu and 1− to 8− esu ranges, respectively. ADP binding then reduces zM to the 7+ to 0.88+ esu range. © 1990, American Chemical Society. All rights reserved.