The study demonstrates that a subtle conformational change at the S-1/S-2 junction plays a critical role in determining the enzymatic activities of smooth muscle myosin.
A monoclonal antibody against subfragment 2 (S-2) of smooth muscle myosin, designated MM-9, was generated and characterized. MM-9 potently inhibited subfragment 1 (S-1) release by papain proteolysis of myosin, suggesting that the epitope of MM-9 is at or very close to the S-1/S-2 junction. The depression of Ca2(+)- and Mg2(+)-ATPase activities of myosin at low ionic strength was significantly reduced by MM-9. MM-9 increased the acto dephosphorylated HMM ATPase activity about 3-fold. On the other hand, the antibody had no effect on the KCl-dependence of viscosity of monomeric myosin. These results suggest that the folding of the myosin rod is not the direct determinant of enzymatic activity, and that the subtle conformational change at the S-1/S-2 junction (head-neck region) plays a critical role in determining enzymatic activities.
Higashihara et al. (1990) studied this question.