Structural and mechanistic similarities of 6-phosphogluconate and β-hydroxyisobutyrate dehydrogcnases reveal a new family of enzymes, the β-hydroxyacid dehydrogenases

J. W. Hawes, E. T. Harper, D. W. Crabb, R. A. Harris

Research output: Contribution to journalArticle

Abstract

The catabolism of valine occurs with the formation of a free branched-chain acid, (S)-β-hydroxyisobutyrate or HIBA which is reversibly oxidized to methylmalonate semialdehyde by a highly specific dehydrogenase (HIBADH or 3-hydroxy-2methylpropanoate: NADoxidoreductase, EC 1.1.1.31). Previous studies placed this enzyme in the now well established short-chain dehydrogenase gene family based on sequence homology and enzymatic properties such as the lack of a metal requirement for catalysis. However, site-directed mutagenesis studies showed that HIBADH differs mechanistically from the short-chain dehydrogenases. We now show that HIBADH shares more significant sequence homology with a separate group of enzymes including 6-phosphogluconate dehydrogenase from numerous species, and several unidentified ORFs of microbial origin. Most of the amino acid residues suggested to be important for substrate-binding and catalysis from X-ray crystal lographic studies of sheep 6-phosphogluconate dehydrogenase are completely conserved in all of these sequences. Furthermore, a conserved lysine residue suggested to be critical to catalysis in 6-phosphogluconate dehydrogenase is now shown by site-directed mutagenesis to be critical to catalysis by HIBADH. This study suggests that 6-phosphogluconate dehydrogenase and HIBADH may share a common evolutionär)' origin and a common enzymatic mechanism. We propose that these homologous proteins represent a distinct family of enzymes most likely consisting of β-hydroxyacid dehydrogenases.

Original languageEnglish (US)
Pages (from-to)A1245
JournalFASEB Journal
Volume10
Issue number6
StatePublished - Dec 1 1996

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Phosphogluconate Dehydrogenase
phosphogluconate dehydrogenase
new family
Catalysis
catalytic activity
Oxidoreductases
Mutagenesis
3-hydroxyisobutyrate dehydrogenase
site-directed mutagenesis
Enzymes
Sequence Homology
enzymes
Site-Directed Mutagenesis
sequence homology
Valine
valine
Open Reading Frames
Lysine
crystals
open reading frames

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

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title = "Structural and mechanistic similarities of 6-phosphogluconate and β-hydroxyisobutyrate dehydrogcnases reveal a new family of enzymes, the β-hydroxyacid dehydrogenases",
abstract = "The catabolism of valine occurs with the formation of a free branched-chain acid, (S)-β-hydroxyisobutyrate or HIBA which is reversibly oxidized to methylmalonate semialdehyde by a highly specific dehydrogenase (HIBADH or 3-hydroxy-2methylpropanoate: NADoxidoreductase, EC 1.1.1.31). Previous studies placed this enzyme in the now well established short-chain dehydrogenase gene family based on sequence homology and enzymatic properties such as the lack of a metal requirement for catalysis. However, site-directed mutagenesis studies showed that HIBADH differs mechanistically from the short-chain dehydrogenases. We now show that HIBADH shares more significant sequence homology with a separate group of enzymes including 6-phosphogluconate dehydrogenase from numerous species, and several unidentified ORFs of microbial origin. Most of the amino acid residues suggested to be important for substrate-binding and catalysis from X-ray crystal lographic studies of sheep 6-phosphogluconate dehydrogenase are completely conserved in all of these sequences. Furthermore, a conserved lysine residue suggested to be critical to catalysis in 6-phosphogluconate dehydrogenase is now shown by site-directed mutagenesis to be critical to catalysis by HIBADH. This study suggests that 6-phosphogluconate dehydrogenase and HIBADH may share a common evolution{\"a}r)' origin and a common enzymatic mechanism. We propose that these homologous proteins represent a distinct family of enzymes most likely consisting of β-hydroxyacid dehydrogenases.",
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AU - Crabb, D. W.

AU - Harris, R. A.

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N2 - The catabolism of valine occurs with the formation of a free branched-chain acid, (S)-β-hydroxyisobutyrate or HIBA which is reversibly oxidized to methylmalonate semialdehyde by a highly specific dehydrogenase (HIBADH or 3-hydroxy-2methylpropanoate: NADoxidoreductase, EC 1.1.1.31). Previous studies placed this enzyme in the now well established short-chain dehydrogenase gene family based on sequence homology and enzymatic properties such as the lack of a metal requirement for catalysis. However, site-directed mutagenesis studies showed that HIBADH differs mechanistically from the short-chain dehydrogenases. We now show that HIBADH shares more significant sequence homology with a separate group of enzymes including 6-phosphogluconate dehydrogenase from numerous species, and several unidentified ORFs of microbial origin. Most of the amino acid residues suggested to be important for substrate-binding and catalysis from X-ray crystal lographic studies of sheep 6-phosphogluconate dehydrogenase are completely conserved in all of these sequences. Furthermore, a conserved lysine residue suggested to be critical to catalysis in 6-phosphogluconate dehydrogenase is now shown by site-directed mutagenesis to be critical to catalysis by HIBADH. This study suggests that 6-phosphogluconate dehydrogenase and HIBADH may share a common evolutionär)' origin and a common enzymatic mechanism. We propose that these homologous proteins represent a distinct family of enzymes most likely consisting of β-hydroxyacid dehydrogenases.

AB - The catabolism of valine occurs with the formation of a free branched-chain acid, (S)-β-hydroxyisobutyrate or HIBA which is reversibly oxidized to methylmalonate semialdehyde by a highly specific dehydrogenase (HIBADH or 3-hydroxy-2methylpropanoate: NADoxidoreductase, EC 1.1.1.31). Previous studies placed this enzyme in the now well established short-chain dehydrogenase gene family based on sequence homology and enzymatic properties such as the lack of a metal requirement for catalysis. However, site-directed mutagenesis studies showed that HIBADH differs mechanistically from the short-chain dehydrogenases. We now show that HIBADH shares more significant sequence homology with a separate group of enzymes including 6-phosphogluconate dehydrogenase from numerous species, and several unidentified ORFs of microbial origin. Most of the amino acid residues suggested to be important for substrate-binding and catalysis from X-ray crystal lographic studies of sheep 6-phosphogluconate dehydrogenase are completely conserved in all of these sequences. Furthermore, a conserved lysine residue suggested to be critical to catalysis in 6-phosphogluconate dehydrogenase is now shown by site-directed mutagenesis to be critical to catalysis by HIBADH. This study suggests that 6-phosphogluconate dehydrogenase and HIBADH may share a common evolutionär)' origin and a common enzymatic mechanism. We propose that these homologous proteins represent a distinct family of enzymes most likely consisting of β-hydroxyacid dehydrogenases.

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