Sensitivity of actomyosin ATPase to calcium and strontium ions. Effect of hybrid troponins.
1. Hybrid or reconstituted troponins were prepared from troponin components of rabbit skeletal muscle and porcine cardiac muscle and their effect on the actomyosin ATPase activity was measured at various concentrations of Ca2+ or Sr2+. The Ca2+ concentration required for half-maximum activation of actomyosin ATPase with troponin containing cardiac troponin I was slightly higher than that with troponin containing skeletal troponin I. The Sr2+ concentration required for half-maximum activation of actomyosin ATPase with troponin containing skeletal troponin C was higher than that with troponin containing cardiac troponin C. 2. Reconstituted cardiac troponin was phosphorylated by cyclic AMP-dependent protein kinase. The Ca2+ sensitivity of actomyosin ATPase with cardiac troponin decreased upon phosphorylation of troponin I; maximum ATPase activity was depressed and the Ca2+ concentration at half-maximum activation increased. On the other hand, phosphorylation of troponin I did not change Sr2+ sensitivity. 3. The inhibitory effect of cardiac troponin I on the actomyosin ATPase activity was neutralized by increasing the amount of brain calmodulin at high Ca2+ and Sr2+ concentrations but not at low concentrations. 4. ATPase activity of actomyosin with a mixture of troponin I and calmodulin was assayed at various concentrations of Ca2+ or Sr2+. The Ca2+ or Sr2+ sensitivity of actomyosin ATPase containing skeletal troponin I was approximately the same as that of actomyosin ATPase containing cardiac troponin I. Phosphorylation of cardiac troponin I did not change the Ca2+ sensitivity of the ATPase. 5. The Ca2+ or Sr2+ concentration required for half-maximum activation of actomyosin ATPase with troponin I-T-calmodulin was higher than that of actomyosin ATPase with the mixture of troponin I and calmodulin. Maximum ATPase activity was lower than that with the mixture of troponin I and calmodulin.
D000205:Actomyosin / Q000378:metabolism*; D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors*; D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002147:Calmodulin / Q000378:metabolism; D004285:Dogs; D009124:Muscle Proteins / Q000378:metabolism*; D009132:Muscles / Q000201:enzymology; D010766:Phosphorylation; D011817:Rabbits; D013324:Strontium / Q000494:pharmacology*; D013552:Swine; D014335:Tropomyosin / Q000378:metabolism; D014336:Troponin / Q000378:metabolism*; D019209:Troponin C; D019210:Troponin I
D002147:Calmodulin; D009124:Muscle Proteins; D014335:Tropomyosin; D014336:Troponin; D019209:Troponin C; D019210:Troponin I; D000205:Actomyosin; D000251:Adenosine Triphosphatases; D002118:Calcium; D013324:Strontium