Mammalian and avian liver phosphoenolpyruvate carboxykinase: Alternate substrates and inhibition by analogues of oxaloacetate

David E. Ash, Frances A. Emig, Soheli A. Chowdhury, Yoshitake Satoh, Vern L. Schramm

Research output: Contribution to journalArticle

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Abstract

Phosphoenolpyruvate carboxykinase from chicken liver mitochondria and rat liver cytosol catalyzes the phosphorylation of α-substituted carboxylic acids such as glycolate, thioglycolate, and DL-β-chlorolactate in reactions with absolute requirements for divalent cation activators. 31P NMR analysis of the reaction products indicates that phosphorylation occurs at the α-position to generate the corresponding O- or S-bridged phosphate monoesters. In addition, the enzymes catalyze the bicarbonate-dependent phosphorylation of hydroxylamine. The chicken liver enzyme also catalyzes the bicarbonate-dependent phosphorylation of fluoride ion. The kcat values for these substrates are 20-1000-fold slower than the kcat for oxaloacetate. Pyruvate and β-hydroxypyruvate are not phosphorylated, since the enzyme does not catalyze the enolization of these compounds. Oxalate, a structural analogue of the enolate of pyruvate, is a competitive inhibitor of phosphoenolpyruvate carboxykinase (Ki of 5 μM) in the direction of phosphoenolpyruvate formation. Oxalate is also an inhibitor of the chicken liver enzyme in the direction of oxaloacetate formation and in the decarboxylation of oxaloacetate. The chicken liver enzyme is inhibited by β-sulfopyruvate, an isoelectronic analogue of oxaloacetate. The extensive homologies between the reactions catalyzed by phosphoenolpyruvate carboxykinase and pyruvate kinase suggest that the divalent cation activators in these reactions may have similar functions. The substrate specificity indicates that phosphoenolpyruvate carboxykinase decarboxylates oxaloacetate to form the enolate of pyruvate which is then phosphorylated by MgGTP on the enzyme.

Original languageEnglish (US)
Pages (from-to)7377-7384
Number of pages8
JournalJournal of Biological Chemistry
Volume265
Issue number13
StatePublished - May 5 1990

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Oxaloacetic Acid
Phosphoenolpyruvate
Liver
Phosphorylation
Chickens
Substrates
Enzymes
Pyruvic Acid
glycolic acid
Oxalates
Divalent Cations
Bicarbonates
Thioglycolates
Hydroxylamine
Decarboxylation
Mitochondria
Pyruvate Kinase
Liver Mitochondrion
Substrate Specificity
Carboxylic Acids

ASJC Scopus subject areas

  • Biochemistry

Cite this

Mammalian and avian liver phosphoenolpyruvate carboxykinase : Alternate substrates and inhibition by analogues of oxaloacetate. / Ash, David E.; Emig, Frances A.; Chowdhury, Soheli A.; Satoh, Yoshitake; Schramm, Vern L.

In: Journal of Biological Chemistry, Vol. 265, No. 13, 05.05.1990, p. 7377-7384.

Research output: Contribution to journalArticle

Ash, David E. ; Emig, Frances A. ; Chowdhury, Soheli A. ; Satoh, Yoshitake ; Schramm, Vern L. / Mammalian and avian liver phosphoenolpyruvate carboxykinase : Alternate substrates and inhibition by analogues of oxaloacetate. In: Journal of Biological Chemistry. 1990 ; Vol. 265, No. 13. pp. 7377-7384.
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