Stereoselectivity of enoyl-CoA hydratase results from preferential activation of one of two bound substrate conformers

Chem Biol. 2002 Nov;9(11):1247-55. doi: 10.1016/s1074-5521(02)00263-6.


Enoyl-CoA hydratase catalyzes the hydration of trans-2-crotonyl-CoA to 3(S)- and 3(R)-hydroxybutyryl-CoA with a stereoselectivity (3(S)/3(R)) of 400,000 to 1. Importantly, Raman spectroscopy reveals that both the s-cis and s-trans conformers of the substrate analog hexadienoyl-CoA are bound to the enzyme, but that only the s-cis conformer is polarized. This selective polarization is an example of ground state strain, indicating the existence of catalytically relevant ground state destabilization arising from the selective complementarity of the enzyme toward the transition state rather than the ground state. Consequently, the stereoselectivity of the enzyme-catalyzed reaction results from the selective activation of one of two bound substrate conformers rather than from selective binding of a single conformer. These findings have important implications for inhibitor design and the role of ground state interactions in enzyme catalysis.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Acyl Coenzyme A / chemistry
  • Acyl Coenzyme A / metabolism
  • Animals
  • Binding Sites
  • Catalysis
  • Crystallography, X-Ray
  • Enoyl-CoA Hydratase / chemistry
  • Enoyl-CoA Hydratase / genetics
  • Enoyl-CoA Hydratase / metabolism*
  • Enzyme Inhibitors / chemistry
  • Molecular Structure
  • Mutation
  • Rats
  • Recombinant Proteins
  • Spectrum Analysis, Raman
  • Stereoisomerism
  • Substrate Specificity


  • Acyl Coenzyme A
  • Enzyme Inhibitors
  • Recombinant Proteins
  • hexadienoyl-coenzyme A
  • Enoyl-CoA Hydratase