Substrate specificity combined with stereopromiscuity in glutathione transferase A4-4-dependent metabolism of 4-hydroxynonenal

Biochemistry. 2010 Feb 23;49(7):1541-8. doi: 10.1021/bi902038u.

Abstract

Conjugation to glutathione (GSH) by glutathione transferase A4-4 (GSTA4-4) is a major route of elimination for the lipid peroxidation product 4-hydroxynonenal (HNE), a toxic compound that contributes to numerous diseases. Both enantiomers of HNE are presumed to be toxic, and GSTA4-4 has negligible stereoselectivity toward them, despite its high catalytic chemospecificity for alkenals. In contrast to the highly flexible, and substrate promiscuous, GSTA1-1 isoform that has poor catalytic efficiency with HNE, GSTA4-4 has been postulated to be a rigid template that is preorganized for HNE metabolism. However, the combination of high substrate chemoselectivity and low substrate stereoselectivity is intriguing. The mechanism by which GSTA4-4 achieves this combination is important, because it must metabolize both enantiomers of HNE to efficiently detoxify the biologically formed mixture. The crystal structures of GSTA4-4 and an engineered variant of GSTA1-1 with high catalytic efficiency toward HNE, cocrystallized with a GSH-HNE conjugate analogue, demonstrate that GSTA4-4 undergoes no enantiospecific induced fit; instead, the active site residue Arg15 is ideally located to interact with the 4-hydroxyl group of either HNE enantiomer. The results reveal an evolutionary strategy for achieving biologically useful stereopromiscuity toward a toxic racemate, concomitant with high catalytic efficiency and substrate specificity toward an endogenously formed toxin.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Aldehydes / chemistry
  • Aldehydes / metabolism*
  • Aldehydes / toxicity*
  • Biocatalysis
  • Catalytic Domain / genetics
  • Crystallography, X-Ray
  • Glutathione Transferase / chemistry*
  • Glutathione Transferase / genetics
  • Glutathione Transferase / metabolism*
  • Humans
  • Ligands
  • Oxidative Stress / genetics
  • Protein Binding
  • Protein Conformation
  • Protein Structure, Secondary / genetics
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Stereoisomerism
  • Substrate Specificity / genetics

Substances

  • Aldehydes
  • Ligands
  • Recombinant Proteins
  • GSTA1 protein, human
  • Glutathione Transferase
  • leukotriene-C4 synthase
  • 4-hydroxy-2-nonenal

Associated data

  • PDB/3IK7
  • PDB/3IK9