Studies on endogenous phosphorylation of chicken erythrocyte membranes. Calcium-dependent phosphorylation of specific proteins.
In this work, endogenous phosphorylation of chicken erythrocyte membranes was investigated. The membrane proteins were rapidly phosphorylated endogenously (half maximum time was 30 s) in the presence of millimolar concentration of Mg2+ under physiological conditions. As an exogenous substrate, protamine was phosphorylated most rapidly of those tested, and histone, casein and bovine serum albumin were rather poor substrates. Cyclic nucleotides had no effect on the endogenous membrane phosphorylation. EGTA inhibited the phosphorylation of a membrane protein having an approximate molecular weight of 43000, and this inhibition was reversed by the addition of a stoichiometric amount of Ca2+. Furthermore, trifluoperazine, an inhibitor of calmodulin, was found to have the same effect as that of EGTA. The phosphorylated 43 kDa protein could be extracted from the membranes under high salt conditions, and was precipitated specifically with anti-actin antibody. These results suggest that the phosphorylation of a peripheral membrane protein (which has an approximate molecular weight of 43000) of chicken erythrocytes by membranous protein kinase(s) depends on Ca2+ and possibly on calmodulin.
D000255:Adenosine Triphosphate / Q000097:blood; D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002645:Chickens; D004910:Erythrocyte Membrane / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism*; D007700:Kinetics; D008565:Membrane Proteins / Q000097:blood* / Q000302:isolation & purification; D008970:Molecular Weight; D010766:Phosphorylation
D008565:Membrane Proteins; D000255:Adenosine Triphosphate; D002118:Calcium