Phosphocretine production coupled to the glycolytic reactions in the cytosol of cardiac cells.
Phosphocreatine production ctalyzed by a cytosolic fraction from cardiac muscle containing all glycolytic enzymes and creatine kinase in a soluble form has been studied in the presence of creatine, adenine nucleotides and different glycolytic intermedites as substrates. Glycolytic depletion of glucose, fructose 1,6bis(phosphate) and phosphoenolpyruvate to lactte was coupled to efficient phosphocreatine production. The molar ratio of phosphocreatine to lactate produced was close to 2.0 when fructose 1,6bis(phosphate) was used as substrate and 1.0 with phosphoenolpyruvate. In these processes the creatine kinase reaction was not the rate-limiting step: themass action ratio of the creatine kinase reaction was very close to its equilibrium value and the maximal rate of the forward creatine kinase reaction exceeded that of glycolytic flux by about 6-fold when fructose 1,6-bis(phosphate) was used as a substrate. Therefore, the creatine kinase raction was continuously in the state of quasi-equilibrium and the efficient syntheses of phosphocreatine observed is a result of constant removal of ADP by the glycolytic system at an almost unchanged level of ATP ([ATP]>>[ADP]), this leading to a continuous shift of the creatine kinase equilibrium position. When phosphocreatine was added initially at concentrations of 5---15 mM the rate of the coupled creatine kinase and glycolytic reactions was very significantly inhibited due to a sharp decrease in the steady-state concentration of ADP. Therefore, under conditions of effective phosphocreatine production in heart mitochondria, which maintain a high phosphocreatine: creatine ratio in the myoplasm in vivo, the glycolytic flux may be suppressed due to limited availability of ADP restricted by the creatine kinase system. The possible physiological role of the control of the glycolytic flux by the creatine kinase system is discussed.
D003402:Creatine Kinase / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism*; D005635:Fructosediphosphates / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006019:Glycolysis; D007700:Kinetics; D008954:Models, Biological; D009206:Myocardium / Q000166:cytology; D010725:Phosphocreatine / Q000096:biosynthesis*; D010728:Phosphoenolpyruvate / Q000378:metabolism; D011770:Pyruvate Kinase / Q000378:metabolism
D005635:Fructosediphosphates; D010725:Phosphocreatine; D010728:Phosphoenolpyruvate; D011770:Pyruvate Kinase; D003402:Creatine Kinase; D005947:Glucose