Isoenzymes of pyruvate dehydrogenase phosphatase. DNA-derived amino acid sequences, expression, and regulation

Boli Huang, Ramadevi Gudi, Pengfei Wu, Robert Harris, Jean Hamilton, Kirill M. Popov

Research output: Contribution to journalArticle

154 Citations (Scopus)

Abstract

Pyruvate dehydrogenase phosphatase (PDP) is one of the few mammalian phosphatases residing within the mitochondrial matrix space. It is responsible for dephosphorylation and reactivation of the pyruvate dehydrogenase complex (PDC) and, by this means, is intimately involved in the regulation of utilization of carbohydrate fuels in mammals. PDP is a dimeric enzyme consisting of catalytic and regulatory subunits. The catalytic subunit of PDP is a Mg2+-dependent enzyme homologous to the cytosolic phosphatases of the 2C family. In the present study, we isolated two cDNAs encoding for mitochondrial phosphatases. The first cDNA is highly homologous to the previously identified cDNA encoding for the catalytic subunit of PDP (PDP1). The second cDNA encodes a previously unknown catalytic subunit of PDP (PDP2). The new phosphatase, expressed as the recombinant protein in Escherichia coli, shows strict substrate specificity toward PDC and does not use phosphorylated branched chain α-ketoacid dehydrogenase as substrate. Like PDP1, PDP2 is a Mg2+-dependent enzyme, but its sensitivity to Mg2+ ions is almost 10-fold lower than that of PDP1. In contrast to PDP1, PDP2 is not regulated by Ca2+ ions. Instead, it is sensitive to the biological polyamine spermine, which, in turn, has no effect on the enzymatic activity of PDP1. Western blot analysis of PDP extracted from mitochondria isolated from liver and skeletal muscle revealed that PDP1 is predominantly expressed in mitochondria from skeletal muscle, whereas PDP2 is much more abundant in the liver rather than muscle mitochondria. Both isoenzymes are expressed in mitochondria from 3T3-L1 adipocytes, but the level of expression of PDP2 is considerably higher. These observations are consistent with previous findings on the enzymatic parameters of PDP in adipose tissue. Thus, our results provide the first evidence that there are at least two isoenzymes of PDP in mammals that are different with respect to tissue distribution and kinetic parameters and, therefore, are likely to be different functionally.

Original languageEnglish
Pages (from-to)17680-17688
Number of pages9
JournalJournal of Biological Chemistry
Volume273
Issue number28
DOIs
StatePublished - Jul 10 1998

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Pyruvate Dehydrogenase (Lipoamide)-Phosphatase
Isoenzymes
Amino Acid Sequence
Amino Acids
Mitochondria
DNA
Phosphoric Monoester Hydrolases
Catalytic Domain
Complementary DNA
Pyruvate Dehydrogenase Complex
Muscle
Mammals
Liver
Skeletal Muscle
Enzymes
3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)
Ions
Tissue
Muscle Mitochondrion
Spermine

ASJC Scopus subject areas

  • Biochemistry

Cite this

Isoenzymes of pyruvate dehydrogenase phosphatase. DNA-derived amino acid sequences, expression, and regulation. / Huang, Boli; Gudi, Ramadevi; Wu, Pengfei; Harris, Robert; Hamilton, Jean; Popov, Kirill M.

In: Journal of Biological Chemistry, Vol. 273, No. 28, 10.07.1998, p. 17680-17688.

Research output: Contribution to journalArticle

Huang, Boli ; Gudi, Ramadevi ; Wu, Pengfei ; Harris, Robert ; Hamilton, Jean ; Popov, Kirill M. / Isoenzymes of pyruvate dehydrogenase phosphatase. DNA-derived amino acid sequences, expression, and regulation. In: Journal of Biological Chemistry. 1998 ; Vol. 273, No. 28. pp. 17680-17688.
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