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

Research output: Contribution to journalArticle

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 (US)
Pages (from-to)36261-36267
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number39
DOIs
StatePublished - Sep 28 2001
Externally publishedYes

Fingerprint

Camphor 5-Monooxygenase
Raman Spectrum Analysis
Cyanides
Raman spectroscopy
Stretching
Derivatives
Fourier Analysis
Isotopes
Liver
Raman scattering
Infrared spectroscopy
Spectrum Analysis
Fourier transforms
Rabbits
Geometry

ASJC Scopus subject areas

  • Biochemistry

Cite this

Elucidation of the Differences between the 430- and 455-nm Absorbing Forms of P450-Isocyanide Adducts by Resonance Raman Spectroscopy. / Tomita, Takeshi; Ogo, Seiji; Egawa, Tsuyoshi; Shimada, Hideo; Okamoto, Noriaki; Imai, Yoshio; Watanabe, Yoshihito; Ishimura, Yuzuru; Kitagawa, Teizo.

In: Journal of Biological Chemistry, Vol. 276, No. 39, 28.09.2001, p. 36261-36267.

Research output: Contribution to journalArticle

Tomita, T, Ogo, S, Egawa, T, Shimada, H, Okamoto, N, Imai, Y, Watanabe, Y, Ishimura, Y & Kitagawa, T 2001, 'Elucidation of the Differences between the 430- and 455-nm Absorbing Forms of P450-Isocyanide Adducts by Resonance Raman Spectroscopy', Journal of Biological Chemistry, vol. 276, no. 39, pp. 36261-36267. https://doi.org/10.1074/jbc.M104932200
Tomita, Takeshi ; Ogo, Seiji ; Egawa, Tsuyoshi ; Shimada, Hideo ; Okamoto, Noriaki ; Imai, Yoshio ; Watanabe, Yoshihito ; Ishimura, Yuzuru ; Kitagawa, Teizo. / Elucidation of the Differences between the 430- and 455-nm Absorbing Forms of P450-Isocyanide Adducts by Resonance Raman Spectroscopy. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 39. pp. 36261-36267.
@article{1b3c69089962403093b22c3c214ef33f,
title = "Elucidation of the Differences between the 430- and 455-nm Absorbing Forms of P450-Isocyanide Adducts by Resonance Raman Spectroscopy",
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.",
author = "Takeshi Tomita and Seiji Ogo and Tsuyoshi Egawa and Hideo Shimada and Noriaki Okamoto and Yoshio Imai and Yoshihito Watanabe and Yuzuru Ishimura and Teizo Kitagawa",
year = "2001",
month = "9",
day = "28",
doi = "10.1074/jbc.M104932200",
language = "English (US)",
volume = "276",
pages = "36261--36267",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "39",

}

TY - JOUR

T1 - Elucidation of the Differences between the 430- and 455-nm Absorbing Forms of P450-Isocyanide Adducts by Resonance Raman Spectroscopy

AU - Tomita, Takeshi

AU - Ogo, Seiji

AU - Egawa, Tsuyoshi

AU - Shimada, Hideo

AU - Okamoto, Noriaki

AU - Imai, Yoshio

AU - Watanabe, Yoshihito

AU - Ishimura, Yuzuru

AU - Kitagawa, Teizo

PY - 2001/9/28

Y1 - 2001/9/28

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0035965195&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035965195&partnerID=8YFLogxK

U2 - 10.1074/jbc.M104932200

DO - 10.1074/jbc.M104932200

M3 - Article

C2 - 11459844

AN - SCOPUS:0035965195

VL - 276

SP - 36261

EP - 36267

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 39

ER -