Resonance Raman study on mutant cytochrome P-450 obtained by site-directed mutagenesis

T. Egawa, Y. Imai, T. Ogura, T. Kitagawa

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

Resonance Raman spectra were observed for the threonine-301 to serine or valine mutant as well as the wild type of rabbit liver microsomal cytochrome P-450 [laurate(ω - 1)-hydroxylase] (P-450(ω - 1), which were prepared through site-directed mutagenesis. The high-spin marker resonance Raman (RR) bands became similarly stronger for all the P-450s examined in the oxidized form upon addition of laurate, and the RR spectra in the higher frequency region of the oxidized, reduced and CO-adduct forms did not distinctly differ among the P-450s examined. Nevertheless, the FeCO stretching mode (νFeCO) of the CO adduct exhibited an upshift for the valine mutant, suggesting positional proximity of Thr-301 to bound CO like Thr-252 of P-450cam, in agreement with the expectation from the sequence analysis. The νFeCO band was shifted to higher frequency upon binding of normal alkyl fatty acids with C10 or longer alkyl chain but little affected by binding of shorter fatty acids.

Original languageEnglish (US)
Pages (from-to)211-216
Number of pages6
JournalBiochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
Volume1040
Issue number2
DOIs
StatePublished - Sep 3 1990

Keywords

  • Carbonmonoxide adduct
  • Cytochrome P-450
  • Laurate(ω - 1)-hydroxylase
  • Resonance Raman

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology

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