Decreased substrate affinity upon alteration of the substrate-docking region in cytochrome P450(BM-3)

Shelley A. Maves, Hyeyeong Yeom, Mark A. McLean, Stephen G. Sligar

Research output: Contribution to journalArticlepeer-review

Abstract

A mutation at the surface of the substrate access channel which dramatically decreases the affinity for some fatty acids in P450(BM-3) was discovered by random mutagenesis. The mutation introduced, proline-25 to glutamine, is in close proximity to the arginine-47 residue thought to be responsible for the initial docking of fatty acid substrates. The P25Q mutant displays an affinity for palmitate which is approximately 100-fold weaker than the wild-type enzyme. In addition to its altered substrate affinity, P25Q also exhibits altered hydroxylation specificity and carbon monoxide recombination kinetics in the substrate-free form.

Original languageEnglish (US)
Pages (from-to)213-218
Number of pages6
JournalFEBS Letters
Volume414
Issue number2
DOIs
StatePublished - Sep 8 1997

Keywords

  • Carbon monoxide geminate kinetics
  • Random mutagenesis
  • Regiospecificity
  • Substrate binding

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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