Effect of phosphorylation by cyclic AMP-dependent protein kinase on the smooth muscle actomyosin Mg2+-ATPase stimulatory activity of fodrin.
Fodrin, a non-erythrocyte spectrin-like protein, has been purified from bovine brain and found to be phosphorylated by the cyclic AMP-dependent protein kinase with a maximal stoichiometry of 1.02 +/- 0.06 mol of phosphate/mol of fodrin dimer (n = 4). This phosphorylation was not affected by the presence of actin and calmodulin. The phosphorylation of fodrin was found to occur exclusively at serine residues on the beta subunit. Two-dimensional thin layer electrophoresis and chromatography of a tryptic digest of phosphorylated fodrin showed one major phosphopeptide and a few minor ones. We have previously reported that nonphosphorylated fodrin is capable of stimulating the smooth muscle actomyosin Mg2+-ATPase by 50-70% under a well-defined set of conditions such as a critical fodrin concentration and an optimal preincubation time (Wang, C., Ngai, P.K., Walsh, M.P., and Wang, J.H. (1987) Biochemistry 24, 1110-1117). We now report that phosphorylation of fodrin completely eliminates this stimulatory effect. However, phosphorylation of fodrin was able to compete with nonphosphorylated fodrin to result in the abolition of the stimulatory effect. Similarly, nonphosphorylated fodrin could overcome the inhibitory effect created by phosphorylated fodrin. The present results support the suggestion that the stimulation of the smooth muscle actomyosin Mg2+-ATPase by fodrin may be a physiological phenomenon and cyclic AMP may serve as a regulator for this effect.
D000818:Animals; D017301:Ca(2+) Mg(2+)-ATPase / Q000032:analysis*; D002118:Calcium / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism* / Q000494:pharmacology; D002417:Cattle; D008840:Microfilament Proteins / Q000378:metabolism* / Q000494:pharmacology; D009130:Muscle, Smooth / Q000201:enzymology*; D010766:Phosphorylation; D011494:Protein Kinases / Q000494:pharmacology*; D013049:Spectrin
D017301:Ca(2+) Mg(2+)-ATPase; D002118:Calcium; D002352:Carrier Proteins; D008840:Microfilament Proteins; D011494:Protein Kinases; D013049:Spectrin