title: Function of M-line-bound creatine kinase as intramyofibrillar ATP regenerator at the receiving end of the phosphorylcreatine shuttle in muscle.
abstract: After 10 wash cycles, 0.8 u.e. of creatine kinase activity remained bound per mg of chicken pectoralis myofibrils which had been freed of soluble creatine kinase, mitochondria, and membranes. The bound creatine kinase is located at the M-band and contributes to the electron density of this sarcomeric structure (Wallimann, T., Pelloni, G.W., Turner, D.C., and Eppenberger, H. M. (1978) Proc. Natl. Acad. Sci. U. S. A. 75, 4296-4300). By measuring the combined actin-activated Mg2+-ATPase and creatine kinase reactions of myofibrils by pH-stat, it was shown that the amount of M-line-bound creatine kinase activity was sufficient to rephosphorylate the ATP hydrolyzed in vitro by the actin-activated Mg2+-ATPase. The amount of M-line-bound creatine kinase and thus the ATP regeneration potential depended on the muscle type. It was higher in fast muscles and lower in slow muscles. Inhibition of myofibrillar creatine kinase or extraction of the M-line-bound enzyme abolished the ATP regeneration potential without affecting ATPase activity. Inhibitors of myokinase, mitochondrial ADP/ATP translocase, and respiration did not affect the ATP regeneration potential or the ATPase. M-line-bound creatine kinase, sufficient to support an ATP turnover rate of 6s-1 per myosin head, seems to have the capacity for the intramyofibrillar regeneration of most or all of the ATP hydrolyzed by the myofibrillar ATPase during muscle contraction. Thus, M-line-bound creatine kinase at the myofibrillar receiving end of the phosphorylcreatine shuttle is of physiological significance.
mesh_terms: D000199:Actins / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D017301:Ca(2+) Mg(2+)-ATPase; D002645:Chickens; D003402:Creatine Kinase / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes; D007700:Kinetics; D009132:Muscles / Q000201:enzymology*; D009210:Myofibrils / Q000201:enzymology*; D010725:Phosphocreatine / Q000378:metabolism*
chemical_list: D000199:Actins; D007527:Isoenzymes; D010725:Phosphocreatine; D000255:Adenosine Triphosphate; D003402:Creatine Kinase; D000251:Adenosine Triphosphatases; D017301:Ca(2+) Mg(2+)-ATPase