Light-chain phosphorylation controls the conformation of vertebrate non-muscle and smooth muscle myosin molecules.

Phosphorylation of the 20,000-molecular weight (Mr) light chains of vertebrate non-muscle (thymus) and smooth muscle (gizzard) myosins regulates the assembly of these myosins into filaments in vitro. At physiological ionic strength and pH, nonphosphorylated smooth muscle and non-muscle myosin filaments are disassembled by stoichiometric levels of MgATP, forming species having sedimentation coefficients of approximately 11S (range 10-12S; myosin monomers in high salt sediment at 6S). When the 20,000 (20K)-Mr light chains on these 11S myosin species are phosphorylated by the light-chain kinase/calmodulin-Ca2+ complex, the inhibitory effect of the light chains on filament formation is removed and the myosins reassemble into filaments which are stable in MgATP. It was originally suggested that the 11S myosin species was a dimer, previously suggested as a building block for smooth muscle and non-muscle myosin filaments. It has since been shown, however, that 11S smooth muscle myosin is monomeric and has a folded conformation rather than the extended shape characteristic of monomeric myosin in high salt. Here we show that 11S non-muscle myosin is also folded and that phosphorylation of the 20K-Mr light chains of both vertebrate non-muscle (thymus) and vertebrate smooth muscle (gizzard) myosins causes these folded 11S molecules to unfold into the conventional extended monomeric form, which is able to assemble into filaments.