Studies with a reconstituted muscle glycolytic system. The rate and extent of creatine phosphorylation by anaerobic glycolysis.
A mixture of purified muscle glycolytic enzymes was reconstituted and the mixture shown to behave in a fashion analogous to that occurring in vivo. Glycolysis leads to ATP production in muscle and results in the phosphorylation of creatine. The extent of this phosphorylation by anaerobic glycolysis was shown to depend to a small extent on the relative proportions of available P(i) and creatine initially, but more importantly on the first step in glycolysis, in this case the enzyme phosphorylase. With less than 0.1% of the phosphorylase in the a form, only about one-third of the creatine was phosphorylated in 30min, whereas with 4% or more of phosphorylase a, 90% of the creatine was phosphorylated within this time. Inclusion of an adenosine triphosphatase decreased the steady-state concentration of phosphocreatine in the system. Calculations of the theoretical concentrations of ADP and AMP showed that phosphorylase b was almost inactive even in the presence of 9mum-AMP, because of ATP inhibition. With phosphorylase a present, glycolysis was able to continue at least until the calculated concentration of MgADP(-) was only 7mum, and AMP in the sub-mumolar range. The relation of these values to measured concentrations of nucleotides and to phosphorylase a percentages in intact muscle is discussed.
D000251:Adenosine Triphosphatases / Q000494:pharmacology; D000693:Anaerobiosis; D000818:Animals; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003401:Creatine / Q000378:metabolism*; D003402:Creatine Kinase / Q000378:metabolism*; D006019:Glycolysis / Q000187:drug effects; D006736:Horses; D066298:In Vitro Techniques; D007700:Kinetics; D009132:Muscles / Q000187:drug effects / Q000201:enzymology*; D006005:Phosphorylases / Q000494:pharmacology; D011817:Rabbits; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D013056:Spectrophotometry, Ultraviolet; D013552:Swine; D013696:Temperature; D013997:Time Factors
D006005:Phosphorylases; D003402:Creatine Kinase; D000251:Adenosine Triphosphatases; D003401:Creatine