Cooperation between the two heads of smooth muscle myosin is essential for full activation of the motor function by phosphorylation.
The motor function of smooth muscle myosin (SmM) is regulated by phosphorylation of the regulatory light chain (RLC) bound to the neck region of the SmM heavy chain. It is generally accepted that unphosphorylated RLC induces interactions between the two heads and between the head and the tail, thus inhibiting the motor activity of SmM, whereas phosphorylation of RLC interrupts those interactions, thus reversing the inhibition and restoring the motor activity to the maximal value. One assumption of this model is that single-headed SmM is fully active regardless of phosphorylation. To re-evaluate this model, we produced a number of SmM constructs with coiled coils of various lengths and examined their structure and regulation. With these constructs we identified the segment in the coiled-coil key for the formation of a stable double-headed structure. In agreement with the current model, we found that the actin-activated ATPase activity of unphosphorylated SmM increased with shortening of the coiled-coil. However, contrary to the current model, we found that the actin-activated ATPase activity of phosphorylated SmM decreased with shortening coiled-coil and only the stable double-headed SmM was fully activated by phosphorylation. These results indicate that single-headed SmM is neither fully active nor fully inhibited. Based on our findings, we propose that cooperation between the two heads is essential, not only for the inhibition of unphosphorylated SmM, but also for the activation of phosphorylated SmM.
School of Life Sciences, University of Science and Technology of China , Hefei, Anhui 230027, China.
Rong-Na
RN
Ma
Katsuhide
K
Mabuchi
Jing
J
Li
Zekuan
Z
Lu
Chih-Lueh Albert
CL
Wang
Xiang-dong
XD
Li
D000818:Animals; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D008854:Microscopy, Electron; D009130:Muscle, Smooth / Q000378:metabolism; D018995:Myosin Heavy Chains / Q000737:chemistry / Q000378:metabolism*; D018994:Myosin Light Chains / Q000737:chemistry / Q000378:metabolism*; D015879:Myosin Subfragments / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation
D018994:Myosin Light Chains; D015879:Myosin Subfragments; D018995:Myosin Heavy Chains