Effect of phosphorylation of myosin light chain by myosin light chain kinase and protein kinase C on conformational change and ATPase activities of human platelet myosin.
Human platelet myosin forms 10S and 6S conformations, and its Ca(2+)- and Mg(2+)-ATPase activities are parallel with the transition between 10S and 6S conformation, as judged by the gel filtration, intrinsic fluorescence, and viscosity methods. The 20,000-dalton myosin light chain (LC20) is phosphorylated by both myosin light chain kinase (MLC kinase) and Ca2+, phospholipid-dependent protein kinase (protein kinase C [PKC]). The phosphorylation (1 mol of phosphate/mol of LC20) by MLC kinase shifts the equilibrium toward the 6S conformation, but that by PKC does not. The prephosphorylation of myosin by PKC prevents the effect of phosphorylation by MLC kinase on actin-activated Mg(2+)-ATPase activity, but not the effect on conformational change. Inhibition of actin-activated ATPase activity by PKC is due to a decreased affinity of myosin for actin, and no change in Vmax is observed. These results suggest that sequential phosphorylation of myosin by both kinases plays an important role in the ATPase activities of human platelet myosin.
Higashihara
M
M
First Department of Internal Medicine, Faculty of Medicine, University of Tokyo, Japan.
Takahata
K
K
Kurokawa
K
K
D000199:Actins / Q000494:pharmacology; D000251:Adenosine Triphosphatases / Q000097:blood*; D001792:Blood Platelets / Q000378:metabolism*; D017301:Ca(2+) Mg(2+)-ATPase / Q000378:metabolism; D000252:Calcium-Transporting ATPases / Q000378:metabolism; D006801:Humans; D009219:Myosin-Light-Chain Kinase / Q000378:metabolism*; D009218:Myosins / Q000097:blood* / Q000737:chemistry; D010766:Phosphorylation; D011189:Potassium Chloride / Q000494:pharmacology; D011487:Protein Conformation; D011493:Protein Kinase C / Q000378:metabolism*; D013050:Spectrometry, Fluorescence
D000199:Actins; D011189:Potassium Chloride; D011493:Protein Kinase C; D009219:Myosin-Light-Chain Kinase; D000251:Adenosine Triphosphatases; D017301:Ca(2+) Mg(2+)-ATPase; D009218:Myosins; D000252:Calcium-Transporting ATPases