Smooth muscle calponin. Inhibition of actomyosin MgATPase and regulation by phosphorylation.
Calponin isolated from chicken gizzard smooth muscle inhibits the actin-activated MgATPase activity of smooth muscle myosin in a reconstituted system composed of contractile and regulatory proteins. ATPase inhibition is not due to inhibition of myosin phosphorylation since, at calponin concentrations sufficient to cause maximal ATPase inhibition, myosin phosphorylation was unaffected. Furthermore, calponin inhibited the actin-activated MgATPase of fully phosphorylated or thiophosphorylated myosin. Although calponin is a Ca2(+)-binding protein, inhibition did not require Ca2+. Furthermore, although calponin also binds to tropomyosin, ATPase inhibition was not dependent on the presence of tropomyosin. Calponin was phosphorylated in vitro by protein kinase C and Ca2+/calmodulin-dependent protein kinase II, but not by cAMP- or cGMP-dependent protein kinases, or myosin light chain kinase. Phosphorylation of calponin by either kinase resulted in loss of its ability to inhibit the actomyosin ATPase. The phosphorylated protein retained calmodulin and tropomyosin binding capabilities, but actin binding was greatly reduced. The calponin-actin interaction, therefore, appears to be responsible for inhibition of the actomyosin ATPase. These observations suggest that calponin may be involved in regulating actin-myosin interaction and, therefore, the contractile state of smooth muscle. Calponin function in turn is regulated by Ca2(+)-dependent phosphorylation.
Winder
S J
SJ
Department of Medical Biochemistry, University of Calgary, Alberta, Canada.
Walsh
M P
MP
D000199:Actins / Q000378:metabolism; D000205:Actomyosin / Q000037:antagonists & inhibitors*; D000818:Animals; D001921:Brain / Q000201:enzymology; D017301:Ca(2+) Mg(2+)-ATPase / Q000037:antagonists & inhibitors*; D002135:Calcium-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002148:Calmodulin-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism; D002417:Cattle; D002645:Chickens; D005895:Gizzard, Avian / Q000378:metabolism; D007700:Kinetics; D008840:Microfilament Proteins; D009130:Muscle, Smooth / Q000378:metabolism*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000302:isolation & purification / Q000378:metabolism; D011494:Protein Kinases / Q000302:isolation & purification / Q000378:metabolism; D000096985:Calponins
D000199:Actins; D000205:Actomyosin; D017301:Ca(2+) Mg(2+)-ATPase; D002135:Calcium-Binding Proteins; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002148:Calmodulin-Binding Proteins; D008840:Microfilament Proteins; D011493:Protein Kinase C; D011494:Protein Kinases; D000096985:Calponins