Pig Liver Pyruvate Carboxylase. The Reaction Pathway for the Carboxylation of Pyruvate

Overview

Journal Biochem J

Specialty Biochemistry

Date 1974 May 1

PMID 4447612

Citations 11

Authors

G B Warren

K F Tipton

Affiliations

Soon will be listed here.

Abstract

1. The reaction pathway for the carboxylation of pyruvate, catalysed by pig liver pyruvate carboxylase, was studied in the presence of saturating concentrations of K(+) and acetyl-CoA. 2. Free Mg(2+) binds to the enzyme in an equilibrium fashion and remains bound during all further catalytic cycles. MgATP(2-) binds next, followed by HCO(3) (-) and then pyruvate. Oxaloacetate is released before the random release, at equilibrium, of P(i) and MgADP(-). 3. This reaction pathway is compared with the double displacement (Ping Pong) mechanisms that have previously been described for pyruvate carboxylases from other sources. The reaction pathway proposed for the pig liver enzyme is superior in that it shows no kinetic inconsistencies and satisfactorily explains the low rate of the ATP[unk][(32)P]P(i) equilibrium exchange reaction. 4. Values are presented for the stability constants of the magnesium complexes of ATP, ADP, acetyl-CoA, P(i), pyruvate and oxaloacetate.

Citing Articles

Parameter Reliability and Understanding Enzyme Function.

McDonald A, Tipton K Molecules. 2022; 27(1).

PMID: 35011495 PMC: 8746786. DOI: 10.3390/molecules27010263.

Leptin reverses diabetes by suppression of the hypothalamic-pituitary-adrenal axis.

Perry R, Zhang X, Zhang D, Kumashiro N, Camporez J, Cline G Nat Med. 2014; 20(7):759-63.

PMID: 24929951 PMC: 4344321. DOI: 10.1038/nm.3579.

Inhibitors of Pyruvate Carboxylase.

Zeczycki T, St Maurice M, Attwood P Open Enzym Inhib J. 2011; 3:8-26.

PMID: 22180764 PMC: 3238542. DOI: 10.2174/1874940201003010008.

Activation and inhibition of pyruvate carboxylase from Rhizobium etli.

Zeczycki T, Menefee A, Jitrapakdee S, Wallace J, Attwood P, St Maurice M Biochemistry. 2011; 50(45):9694-707.

PMID: 21958066 PMC: 3210919. DOI: 10.1021/bi201276r.

Factors that influence the translocation of the N-carboxybiotin moiety between the two sub-sites of pyruvate carboxylase.

Goodall G, Baldwin G, Wallace J, Keech D Biochem J. 1981; 199(3):603-9.

PMID: 7340821 PMC: 1163416. DOI: 10.1042/bj1990603.

References

McClure W . A kinetic analysis of coupled enzyme assays. Biochemistry. 1969; 8(7):2782-6. DOI: 10.1021/bi00835a014. View

Keech B, Barritt G . Allosteric activation of sheep kidney pyruvate carboxylase by the magnesium ion (Mg2+) and the magnesium adenosine triphosphate ion (MgATP2-). J Biol Chem. 1967; 242(9):1983-7. View

Barden R, Fung C, UTTER M, Scrutton M . Pyruvate carboxylase from chicken liver. Steady state kinetic studies indicate a "two-site" ping-pong mechanism. J Biol Chem. 1972; 247(4):1323-33. View

Frieden C . Glutamic dehydrogenase. III. The order of substrate addition in the enzymatic reaction. J Biol Chem. 1959; 234:2891-6. View

MELCHIOR N, MELCHIOR J . The role of complex metal ions in the yeast hexokinase reaction. J Biol Chem. 1958; 231(2):609-23. View

Mildvan A, Scrutton M . Pyruvate carboxylase. X. The demonstration of direct coordination of pyruvate and alpha-ketobutyrate by the bound manganese and the formation of enzyme-metal-substrate bridge complexes. Biochemistry. 1967; 6(10):2978-94. DOI: 10.1021/bi00862a003. View

Dixon M . The determination of enzyme inhibitor constants. Biochem J. 1953; 55(1):170-1. PMC: 1269152. DOI: 10.1042/bj0550170. View

Mildvan A, Scrutton M, UTTER M . Pyruvate carboxylase. VII. A possible role for tightly bound manganese. J Biol Chem. 1966; 241(15):3488-98. View

Burton K . Formation constants for the complexes of adenosine di- or tri-phosphate with magnesium or calcium ions. Biochem J. 1959; 71(2):388-95. PMC: 1196802. DOI: 10.1042/bj0710388. View

10.

Duggleby R, Dennis D . Nicotinamide adenine dinucleotide-specific isocitrate dehydrogenase from a higher plant. The requirement for free and metal-complexed isocitrate. J Biol Chem. 1970; 245(15):3745-50. View

11.

Warren G, Tipton K . Pig liver pyruvate carboxylase. Purification, properties and cation specificity. Biochem J. 1974; 139(2):297-310. PMC: 1166285. DOI: 10.1042/bj1390297. View

12.

Scrutton M, Mildvan A . Pyruvate carboxylase: nuclear magnetic resonance studies of the enzyme-manganese-oxalacetate and enzyme-manganese-pyruvate bridge complexes. Arch Biochem Biophys. 1970; 140(1):131-51. DOI: 10.1016/0003-9861(70)90017-2. View

13.

McClure W, LARDY H, Wagner M, Cleland W . Rat liver pyruvate carboxylase. II. Kinetic studies of the forward reaction. J Biol Chem. 1971; 246(11):3579-83. View

14.

Cazzulo J, Sundaram T, Kornberg H . Properties and regulation of pyruvate carboxylase from Bacillus stearothermophilus. Proc R Soc Lond B Biol Sci. 1970; 176(1042):1-19. DOI: 10.1098/rspb.1970.0030. View

15.

OSullivan W, Perrin D . THE STABILITY CONSTANTS OF METAL-ADENINE NUCLEOTIDE COMPLEXES. Biochemistry. 1964; 3:18-26. DOI: 10.1021/bi00889a005. View

16.

Cazzulo J, Stoppani A . Effects of adenosine phosphates and nicotinamide nucleotides on pyruvate carboxylase from baker's yeast. Biochem J. 1969; 112(5):755-62. PMC: 1187780. DOI: 10.1042/bj1120755. View

17.

Blair J . A reinterpretation of the kinetics of pyruvate carboxylase. FEBS Lett. 1969; 2(4):245-247. DOI: 10.1016/0014-5793(69)80032-3. View

18.

McClure W, LARDY H, Kneifel H . Rat liver pyruvate carboxylase. I. Preparation, properties, and cation specificity. J Biol Chem. 1971; 246(11):3569-78. View

19.

Feir H, Suzuki I . Pyruvate carboxylase of Aspergillus niger: kinetic study of a biotin-containing carboxylase. Can J Biochem. 1969; 47(7):697-710. DOI: 10.1139/o69-107. View

20.

Seufert D, Herlemann E, Albrecht E, SEUBERT W . On the mechanism of gluconeogenesis and its regulation. VII. Purification and properties of pyruvate carboxylase from rat liver. Hoppe Seylers Z Physiol Chem. 1971; 352(3):459-78. DOI: 10.1515/bchm2.1971.352.1.459. View