Myosin Light Chain (Ser-1), Phosphospecific Antibody

Both smooth muscle and nonmuscle myosin II activity is regulated by the phosphorylation state of the myosin regulatory light chain (MLC, MRLC, MLC20, Myl9) . Phosphorylation of MLC at Thr-18 and Ser-19 activates myosin II motor activity and increases myosin filament stability. This activation has important roles in various cell motile processes. By contrast, other phosphorylation sites on MLC may inhibit myosin II activity. PKC phosphorylates Ser-1/Ser-2 and Thr-9 in MLC, and this phosphorylation decreases activated myosin II interaction with actin, as well as inhibits MLC interaction with the activation site kinase, myosin light-chain kinase. The Ser-1/Ser-2 region may be the major inhibitory site since Ser-1 is phosphorylated during PDGF-induced stress fiber disassembly and expression of unphosphorylatable MLC20 at the Ser-1/Ser-2 site suppresses this disassembly. Thus, inhibition of myosin II activity through phosphorylation of Ser-1/Ser-2 may have important roles in growth factor-induced reorganization of actomyosin filaments.

Phospho-MLC (Ser-1) synthetic peptide (coupled to KLH) corresponding to amino acid residues surrounding Ser-1 in bovine myosin regulatory light chain 9 (Myl9) .

The antibody detects a 20 kDa* band corresponding to the molecular weight of myosin light chain in human A431 treated with EGF or calyculin A. In addition, the antibody detects the 20 kDa myosin light chain in mouse C2C12, rat PC12, and rabbit fibroblasts treated with calyculin A. This peptide sequence is highly conserved in human, rat, and mouse Myl9, and in other smooth muscle and non-muscle MLCs.