Chemical modification of lysine and arginine residues in the myosin regulatory light chain inhibits phosphorylation.
The contribution of lysine and arginine residues to the substrate specificity of the myosin light-chain kinase has been studied using chemically modified myosin light chains. Succinylation or maleylation of the myosin light chains caused complete inhibition of their phosphorylation. Modification of 50% of the lysine residues resulted in 90% inhibition of phosphorylation and this was accompanied by a 25-fold increase in the apparent Km. In contrast, phosphorylation of the myosin light chains by the cAMP-dependent protein kinase was relatively insensitive to lysine modification, with only a 15% reduction in phosphorylation following succinylation of 50% of the lysine residues. Treatment with either cyclohexane-1,2-dione or camphorquinone-10-sulfonic acid resulted in between 90 and 98% inhibition of myosin light-chain phosphorylation. These reagents caused modification of both lysine and arginine residues, and accordingly only part of the inhibition can be attributed to arginine modification. Modification of all of the cysteine and methionine residues caused only a 40% inhibition of phosphorylation. The results of this study support the concept that lysine and arginine residues act as essential specificity determinants for the myosin light-chain kinase in protein substrates.
Pearson
R B
RB
Kemp
B E
BE
D000818:Animals; D001120:Arginine; D002164:Camphor / Q000031:analogs & derivatives* / Q000494:pharmacology; D002645:Chickens; D005663:Furans / Q000494:pharmacology*; D005895:Gizzard, Avian / Q000378:metabolism; D007700:Kinetics; D008239:Lysine; D008299:Maleic Anhydrides / Q000494:pharmacology*; D009130:Muscle, Smooth / Q000378:metabolism*; D015879:Myosin Subfragments; D009219:Myosin-Light-Chain Kinase; D009218:Myosins / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism*; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors
D005663:Furans; D008299:Maleic Anhydrides; D015879:Myosin Subfragments; D010446:Peptide Fragments; D047428:Protein Kinase Inhibitors; C023633:camphorquinone-10-sulfonic acid; D002164:Camphor; D001120:Arginine; D009219:Myosin-Light-Chain Kinase; D009218:Myosins; D008239:Lysine