The structure of human cytochrome P450 2C9 complexed with flurbiprofen at 2.0-A resolution

J Biol Chem. 2004 Aug 20;279(34):35630-7. doi: 10.1074/jbc.M405427200. Epub 2004 Jun 4.

Abstract

The structure of human P450 2C9 complexed with flurbiprofen was determined to 2.0 A by x-ray crystallography. In contrast to other structurally characterized P450 2C enzymes, 2C5, 2C8, and a 2C9 chimera, the native catalytic domain of P450 2C9 differs significantly in the conformation of the helix F to helix G region and exhibits an extra turn at the N terminus of helix A. In addition, a distinct conformation of the helix B to helix C region allows Arg-108 to hydrogen bond with Asp-293 and Asn-289 on helix I and to interact directly with the carboxylate of flurbiprofen. These interactions position the substrate for regioselective oxidation in a relatively large active site cavity and are likely to account for the high catalytic efficiency exhibited by P450 2C9 for the regioselective oxidation of several anionic non-steroidal anti-inflammatory drugs. The structure provides a basis for interpretation of a number of observations regarding the substrate selectivity of P450 2C9 and the observed effects of mutations on 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

  • Amino Acid Sequence
  • Aryl Hydrocarbon Hydroxylases / chemistry*
  • Aryl Hydrocarbon Hydroxylases / genetics
  • Aryl Hydrocarbon Hydroxylases / metabolism
  • Binding Sites / genetics
  • Catalytic Domain
  • Cytochrome P-450 CYP2C9
  • Flurbiprofen / chemistry*
  • Flurbiprofen / metabolism
  • Humans
  • Molecular Sequence Data
  • Mutation
  • Protein Binding
  • Protein Conformation
  • Substrate Specificity

Substances

  • Flurbiprofen
  • CYP2C9 protein, human
  • Cytochrome P-450 CYP2C9
  • Aryl Hydrocarbon Hydroxylases

Associated data

  • PDB/1R9O