Sequestration of the active site by interdomain shifting: Crystallographic and spectroscopic evidence for distinct conformations of L-3-hydroxyacyl-CoA dehydrogenase

Joseph J. Barycki, Laurie K. O'Brien, Arnold W. Strauss, Leonard J. Banaszak

Research output: Contribution to journalArticle

52 Scopus citations

Abstract

L-3-Hydroxyacyl-CoA dehydrogenase reversibly catalyzes the conversion of L-3-hydroxyacyl-CoA to 3-ketoacyl-CoA concomitant with the reduction of NAD+ to NADH as part of the β-oxidation spiral. In this report, crystal structures have been solved for the apoenzyme, binary complexes of the enzyme with reduced cofactor or 3-hydroxybutyryl-CoA substrate, and an abortive ternary complex of the enzyme with NAD+ and acetoacetyl-CoA. The models illustrate positioning of cofactor and substrate within the active site of the enzyme. Comparison of these structures with the previous model of the enzyme-NAD+ complex reveals that although significant shifting of the NAD+-binding domain relative to the C-terminal domain occurs in the ternary and substrate-bound complexes, there are few differences between the apoenzyme and cofactor-bound complexes. Analysis of these models clarifies the role of key amino acids implicated in catalysis and highlights additional critical residues. Furthermore, a novel charge transfer complex has been identified in the course of abortive ternary complex formation, and its characterization provides additional insight into aspects of the catalytic mechanism of L-3-hydroxyacyl-CoA dehydrogenase.

Original languageEnglish (US)
Pages (from-to)27186-27196
Number of pages11
JournalJournal of Biological Chemistry
Volume275
Issue number35
DOIs
StatePublished - Sep 1 2000

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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