Elucidation of the Differences between the 430- and 455-nm Absorbing Forms of P450-Isocyanide Adducts by Resonance Raman Spectroscopy

Takeshi Tomita, Seiji Ogo, Tsuyoshi Egawa, Hideo Shimada, Noriaki Okamoto, Yoshio Imai, Yoshihito Watanabe, Yuzuru Ishimura, Teizo Kitagawa

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8 Citations (Scopus)

Abstract

Alkylisocyanide adducts of microsomal P450 exist in two interconvertible forms, each giving the Soret maximum around 430 or 455 nm. This is demonstrated with a rabbit liver P450 2B4. Resonance Raman spectra of the 430- and 455-nm forms were examined for typical P450s of the two types as well as for P450 2B4 because the 430-nm form of P450 2B4 is liable to change into P420. P450cam and P450nor were selected as a model of the 430- and 455-nm forms, respectively. For the n-butyl isocyanide (CNBu) adduct, the Fe(II)-CNBu stretching band was observed for the first time at 480/467 cm-1 for P450cam and at 471/459 cm-1 for P450nor with their 12CNBu/ 13CNBu derivatives. For P450cam, but not P450nor, other 13C isotope-sensitive bands were observed at 412/402, 844/835, and 940/926 cm-1. The C-N stretching mode was identified by Fourier transform IR spectroscopy at 2116/2080 cm-1 for P450cam and at 2148/ 2108 cm-1 for P450nor for the 12C/13C derivatives. These findings suggest that the binding geometry of isocyanide differs between the two forms-bent and linear structures for P450cam-CNBu and P450nor-CNBu, respectively. In contrast, in the ferric state, the Raman 13C isotopic frequency shifts, and the IR C-N stretching frequencies (2213/2170 and 2215/2172 cm-1) were similar between P450cam and P450nor, suggesting similar bent structures for both.

Original languageEnglish
Pages (from-to)36261-36267
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number39
DOIs
Publication statusPublished - Sep 28 2001
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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