A SPECIFIC AMINO-ACID-SEQUENCE AT THE HEAD-ROD JUNCTION IS NOT CRITICAL FOR THE PHOSPHORYLATION-DEPENDENT REGULATION OF SMOOTH-MUSCLE MYOSIN

It has been suggested that the structure at the head-rod junction of s mooth muscle myosin is important for the phosphorylation-mediated regu lation of myosin motor activity. To investigate whether a specific ami no acid sequence at the head-rod junction is critical for the regulati on, three smooth muscle myosin mutants in which the sequence at the N- terminal end of S2 is deleted to various extents were expressed in Sf9 cells; 28, 56, and 84 amino acid residues, respectively, at the posit ion immediately C-terminal to the invariant proline (Pro849) were dele ted, and the S1 domain was directly linked to the downstream sequence of the rod. The mutant myosins were expressed, purified, and biochemic ally characterized. All three myosin mutants showed a stable double-he aded structure based upon electron microscopic observation. Both the a ctin-activated ATPase activity and the actin translocating activity of the mutants were completely regulated by the phosphorylation of the r egulatory light chain. The actin sliding velocity of the three mutant myosins was the same as the wild-type recombinant myosin. These result s indicate that a specific amino acid sequence at the head-rod junctio n is not required for the regulation of smooth muscle myosin. The resu lts also suggest that there is no functionally important interaction b etween the regulatory light chain and the heavy chain at the head-rod junction.