Pig heart short chain L-3-hydroxyacyl-CoA dehydrogenase revisited: Sequence analysis and crystal structure determination

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

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Short chain L-3-hydroxyacyl CoA dehydrogenase (SCHAD) is a soluble dimeric enzyme critical for oxidative metabolism of fatty acids. Its primary sequence has been reported to be conserved across numerous tissues and species with the notable exception of the pig heart homologue. Preliminary efforts to solve the crystal structure of the dimeric pig heart SCHAD suggested the unprecedented occurrence of three enzyme subunits within the asymmetric unit, a phenomenon that was thought to have hampered refinement of the initial chain tracing. The recently solved crystal coordinates of human heart SCHAD facilitated a molecular replacement solution to the pig heart SCHAD data. Refinement of the model, in conjunction with the nucleotide sequence for pig heart SCHAD determined in this paper, has demonstrated that the previously published pig heart SCHAD sequence was incorrect. Presented here are the corrected amino acid sequence and the high resolution crystal structure determined for pig heart SCHAD complexed with its NAD+ cofactor (2.8 Å; R(cryst) = 22.4%, R(free) = 28.8%). In addition, the peculiar phenomenon of a dimeric enzyme crystallizing with three subunits contained in the asymmetric unit is described.

Original languageEnglish (US)
Pages (from-to)2010-2018
Number of pages9
JournalProtein Science
Volume8
Issue number10
DOIs
StatePublished - Jan 1 1999

Fingerprint

3-Hydroxyacyl-CoA Dehydrogenase
Sequence Analysis
Swine
Crystal structure
Enzymes
Metabolism
NAD
Amino Acid Sequence
Fatty Acids
Nucleotides
Tissue
Amino Acids
Crystals

Keywords

  • Dehydrogenase
  • Primary sequence
  • SCHAD
  • X-ray structure
  • β-oxidation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Pig heart short chain L-3-hydroxyacyl-CoA dehydrogenase revisited : Sequence analysis and crystal structure determination. / Barycki, Joseph J.; O'Brien, Laurie K.; Birktoft, Jens J.; Strauss, Arnold W.; Banaszak, Leonard J.

In: Protein Science, Vol. 8, No. 10, 01.01.1999, p. 2010-2018.

Research output: Contribution to journalArticle

Barycki, Joseph J. ; O'Brien, Laurie K. ; Birktoft, Jens J. ; Strauss, Arnold W. ; Banaszak, Leonard J. / Pig heart short chain L-3-hydroxyacyl-CoA dehydrogenase revisited : Sequence analysis and crystal structure determination. In: Protein Science. 1999 ; Vol. 8, No. 10. pp. 2010-2018.
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