Phosphorylation of smooth muscle myosin by type II Ca2+/calmodulin-dependent protein kinase.
Brain type II Ca2+/calmodulin-dependent protein kinase was found to phosphorylate smooth muscle myosin, incorporating maximally approximately 2 mol of phosphoryl per mol of myosin, exclusively on the 20,000 dalton light chain subunit. After maximal phosphorylation of myosin or the isolated 20,000 dalton light chain subunit by myosin light chain kinase, the addition of type II Ca2+/calmodulin-dependent protein kinase led to no further incorporation indicating the two kinases phosphorylated a common site. This conclusion was supported by two dimensional mapping of tryptic digests of myosin phosphorylated by the two kinases. By phosphoamino acid analysis the phosphorylated residue was identified as a serine. The phosphorylation by type II Ca2+/calmodulin-dependent protein kinase of myosin resulted in enhancement of its actin-activated Mg2(+)-ATPase activity. Taken together, these data strongly support the conclusion that type II Ca2+/calmodulin-dependent protein kinase phosphorylates the same amino acid residue on the 20,000 dalton light chain subunit of smooth muscle myosin as is phosphorylated by myosin light chain kinase and suggest an alternative mechanism for the regulation of actin-myosin interaction.
Edelman
A M
AM
Department of Pharmacology and Therapeutics, School of Medicine and Biomedical Sciences, State University of New York, Buffalo 14214.
Lin
W H
WH
Osterhout
D J
DJ
Bennett
M K
MK
Kennedy
M B
MB
Krebs
E G
EG
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001345:Autoradiography; D001921:Brain / Q000201:enzymology; D017301:Ca(2+) Mg(2+)-ATPase / Q000378:metabolism*; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel; D009130:Muscle, Smooth / Q000201:enzymology*; D009218:Myosins / Q000378:metabolism*; D010449:Peptide Mapping; D010759:Phosphorus Isotopes; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*
D000596:Amino Acids; D010759:Phosphorus Isotopes; D011494:Protein Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D017301:Ca(2+) Mg(2+)-ATPase; D009218:Myosins