Myokinase and contractile function of glycerinated muscle fibers.
Glycerinated rabbit psoas muscle fibers containing native CPK, ATPase, and myokinase activities were used and isometric contraction and relaxation responses to either ADP or ATP + CP or to ATP alone in the presence and absence of P1, P5-di(adenosine-5'-pentaphosphate), a myokinase inhibitor, were compared. In previous (14) work it was shown that CP generated more efficient and faster contraction and relaxation of glycerinated muscle fibers than ATP. The present work deals with the role of myokinase in the differential response of fibers to CP and ATP. Inhibition of the myokinase activity of these fibers caused slight diminution of the rate of contraction at physiological concentrations of ATP. Uninhibited fibers were not able to reach maximum contraction, because the tension began to drop gradually even in the presence of Ca2+. Addition of Ap5A permitted maximum contraction and the ability to stay at the contracted state. In the case of CP + adenosine nucleotides (ATP or ADP), myokinase activity decreased the rate of tension development which was statistically significant after 5-7 sec of contraction. Thus, a higher tension was obtainable when myokinase was inhibited. At high concentration of adenine nucleotides (greater than 2 mM) and in the absence of Ap5A, not only the maximum tension never was reached, but a spontaneous drop in tension was observed before addition of EGTA, as was seen with ATP alone. Relaxation was faster and more complete in the presence of uninhibited myokinase activity except that the ADP was low (125 mM). These observations provide further evidence for a close functional interaction of these three enzymes in the mechanism of contraction and relaxation, giving further support to the notion of the creatine-phosphocreatine energy shuttle.
D000227:Adenine Nucleotides / Q000494:pharmacology*; D000244:Adenosine Diphosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000263:Adenylate Kinase / Q000502:physiology*; D000818:Animals; D003402:Creatine Kinase / Q000502:physiology*; D004139:Dinitrofluorobenzene / Q000494:pharmacology; D015226:Dinucleoside Phosphates; D004533:Egtazic Acid / Q000494:pharmacology; D005990:Glycerol / Q000494:pharmacology; D007866:Leg; D009119:Muscle Contraction / Q000187:drug effects*; D009132:Muscles / Q000201:enzymology*; D010725:Phosphocreatine / Q000494:pharmacology; D010770:Phosphotransferases / Q000502:physiology*; D011817:Rabbits; D013997:Time Factors
D000227:Adenine Nucleotides; D015226:Dinucleoside Phosphates; D010725:Phosphocreatine; C012275:P(1),P(5)-di(adenosine-5'-)pentaphosphate; D004533:Egtazic Acid; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D004139:Dinitrofluorobenzene; D010770:Phosphotransferases; D003402:Creatine Kinase; D000263:Adenylate Kinase; D005990:Glycerol