Decarboxylation of oxalacetate to pyruvate by purified avian liver phosphoenolpyruvate carboxykinase.
Phosphoenolpyruvate carboxykinase, which has been isolated from chicken liver mitochondria in essentially homogenous form, carries out the irreversible decarboxylation of oxalacetate to pyruvate in the presence of catalytic amounts of GDP or IDP, as well as the reversible decarboxylation of oxalacetate to phosphoenolpyruvate in the presence of substrate amounts of GTP or ITP. The pyruvate- and phosphoenolpyruvate-forming reactions are similar in their nucleoside specificity and appear to be carried out by the same protein. However, the two activities vary markedly in their response to added metal ions and sulfhydryl reagents. Phosphoenolpyruvate formation is completely dependent on the presence of a divalent metal ion, with Mn2+ the most effective species. This reaction is also stimulated by sulfhydryl reagents such as 2-mercaptoethanol. In contrast, the pyruvate-forming reaction is strongly inhibited by divalent metal ions, including Mn2+, and also by moderate concentrations of sulfhydryl reagents. These observations and the demonstration that pyruvate kinase-like activity is very low or absent make it unlikely that pyruvate formation proceeds via phosphoenolpyruvate as an intermediate. Although the pyruvate-forming reaction is inhibited by added metal ions, the reaction is also inhibited by metal-chelating agents such as 8-hydroxyquinoline and o-phenanthroline, suggesting that the reaction is dependent on the presence of a metal ion. It has not been possible, however, to demonstrate that the enzyme is a metalloprotein.
D015082:2,2'-Dipyridyl / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002645:Chickens; D004789:Enzyme Activation / Q000187:drug effects; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D008623:Mercaptoethanol / Q000494:pharmacology; D010071:Oxaloacetates / Q000378:metabolism; D010618:Phenanthrolines / Q000494:pharmacology; D010729:Phosphoenolpyruvate Carboxykinase (GTP) / Q000302:isolation & purification / Q000378:metabolism*; D011773:Pyruvates / Q000378:metabolism; D011804:Quinolines / Q000494:pharmacology; D012265:Ribonucleotides
D010071:Oxaloacetates; D010618:Phenanthrolines; D011773:Pyruvates; D011804:Quinolines; D012265:Ribonucleotides; D008345:Manganese; D015082:2,2'-Dipyridyl; D008623:Mercaptoethanol; D010729:Phosphoenolpyruvate Carboxykinase (GTP); D008274:Magnesium; D002118:Calcium