Calmodulin-dependent spectrin kinase activity in human erythrocytes.

Membrane protein phosphorylation has been studied in intact human erythrocytes and in resealed erythrocyte ghosts by measuring the incorporation of 32P into band 2 of spectrin. alpha-Adrenergic agonists and Ca+2 stimulate 32P-phosphate incorporation, an effect inhibited by trifluoperazine and diminished in resealed ghosts depleted of calmodulin. Ghosts prepared with endogenous calmodulin or resealed around purified calmodulin exhibit norepinephrine- and Ca+2-stimulated phosphorylation only in the presence of [gamma-32P]-ATP. Ghosts resealed with or without calmodulin in the presence of unlabelled ATP show no net gain or loss of 32P in membrane proteins when exposed to norepinephrine or calcium stimulation. These observations suggest that calcium and norepinephrine stimulation of membrane protein phosphorylation is mediated by calmodulin-dependent spectrin kinase activity, rather than by increased turnover by spectrin ATPase or by inhibition of phosphospectrin phosphatase.