Open AccessCalcium- and Calmodulin-Regulated Phosphorylation of Soluble and Membrane Proteins from Corn Coleoptiles
Author(s) -
K. Veluthambi,
B. W. Poovaiah
Publication year - 1984
Publication title -
plant physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.554
H-Index - 312
eISSN - 1532-2548
pISSN - 0032-0889
DOI - 10.1104/pp.76.2.359
Subject(s) - calmodulin , phosphorylation , trifluoperazine , coleoptile , protein phosphorylation , phosphatase , biochemistry , phosphoprotein , calcium , chemistry , egta , biology , protein kinase a , enzyme , organic chemistry
In vitro phosphorylation of several membrane polypeptides and soluble polypeptides from corn (Zea mays var. Patriot) coleoptiles was promoted by adding Ca(2+). Ca(2+)-promoted phosphorylation of the membrane polypeptides was further increased in the presence of calmodulin. Both Ca(2+)-stimulated and Ca(2+)- and calmodulin-stimulated phosphorylations of membrane polypeptides were inhibited by chlorpromazine, a calmodulin antagonist. Ca(2+)-stimulated phosphorylation of soluble polypeptides increased with increasing Ca(2+) concentration. The calmodulin antagonists chlorpromazine and trifluoperazine inhibited the Ca(2+)-promoted phosphorylation of soluble polypeptides. Added calmodulin promoted the Ca(2+)-dependent phosphorylation of a 98 kilodaltons polypeptide. Both Ca(2+)-dependent and Ca(2+)-independent phosphorylations required Mg(2+) at an optimal concentration of 5 to 10 millimolar. Cyclic AMP was found to have no stimulatory effect on protein phosphorylation. Sodium molybdate, an inhibitor of protein phosphatase, increased the net phosphorylation of several polypeptides. Rapid loss of radioactivity from the phosphorylated polypeptides following incubation in unlabeled ATP indicated the presence of phosphoprotein phosphatase activity.