Resonance Raman detection of the hydroperoxo intermediate in the cytochrome P450 enzymatic cycle

Piotr J. Mak; Ilia G. Denisov; Doreen Victoria; Thomas M. Makris; Tianjing Deng; Stephen G. Sligar; James R. Kincaid

doi:10.1021/ja071426h

Resonance Raman detection of the hydroperoxo intermediate in the cytochrome P450 enzymatic cycle

Piotr J. Mak, Ilia G. Denisov, Doreen Victoria, Thomas M. Makris, Tianjing Deng, Stephen G. Sligar, James R. Kincaid

Research output: Contribution to journal › Article › peer-review

Abstract

The resonance Raman spectra of the hydroperoxo complex of camphor-bound CYP101 have been obtained by cryoradiolytic reduction of the oxygenated ferrous form that had been rapidly frozen in water/glycerol frozen solution; EPR spectroscopy was employed to confirm the identity of the trapped intermediate. The ν(O-O) mode, appearing at 799 cm^-1, is observed for the first time in a peroxo-heme adduct. It is assigned unambiguously by employing isotopomeric mixtures of oxygen gas containing 50% ¹⁶O¹⁸O, confirming the presence of an intact O-O fragment. The ν(Fe-O) mode is observed at 559 cm^-1 (H₂O). Furthermore, both modes shift down by 3 cm^-1, documenting the formulation as a hydroperoxo complex, in agreement with EPR data.

Original language	English (US)
Pages (from-to)	6382-6383
Number of pages	2
Journal	Journal of the American Chemical Society
Volume	129
Issue number	20
DOIs	https://doi.org/10.1021/ja071426h
State	Published - May 23 2007
Externally published	Yes

ASJC Scopus subject areas

Catalysis
General Chemistry
Biochemistry
Colloid and Surface Chemistry

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10.1021/ja071426h

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title = "Resonance Raman detection of the hydroperoxo intermediate in the cytochrome P450 enzymatic cycle",

abstract = "The resonance Raman spectra of the hydroperoxo complex of camphor-bound CYP101 have been obtained by cryoradiolytic reduction of the oxygenated ferrous form that had been rapidly frozen in water/glycerol frozen solution; EPR spectroscopy was employed to confirm the identity of the trapped intermediate. The ν(O-O) mode, appearing at 799 cm-1, is observed for the first time in a peroxo-heme adduct. It is assigned unambiguously by employing isotopomeric mixtures of oxygen gas containing 50% 16O18O, confirming the presence of an intact O-O fragment. The ν(Fe-O) mode is observed at 559 cm-1 (H2O). Furthermore, both modes shift down by 3 cm-1, documenting the formulation as a hydroperoxo complex, in agreement with EPR data.",

author = "Mak, {Piotr J.} and Denisov, {Ilia G.} and Doreen Victoria and Makris, {Thomas M.} and Tianjing Deng and Sligar, {Stephen G.} and Kincaid, {James R.}",

year = "2007",

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language = "English (US)",

volume = "129",

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journal = "Journal of the American Chemical Society",

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T1 - Resonance Raman detection of the hydroperoxo intermediate in the cytochrome P450 enzymatic cycle

AU - Mak, Piotr J.

AU - Denisov, Ilia G.

AU - Victoria, Doreen

AU - Makris, Thomas M.

AU - Deng, Tianjing

AU - Sligar, Stephen G.

AU - Kincaid, James R.

PY - 2007/5/23

Y1 - 2007/5/23

N2 - The resonance Raman spectra of the hydroperoxo complex of camphor-bound CYP101 have been obtained by cryoradiolytic reduction of the oxygenated ferrous form that had been rapidly frozen in water/glycerol frozen solution; EPR spectroscopy was employed to confirm the identity of the trapped intermediate. The ν(O-O) mode, appearing at 799 cm-1, is observed for the first time in a peroxo-heme adduct. It is assigned unambiguously by employing isotopomeric mixtures of oxygen gas containing 50% 16O18O, confirming the presence of an intact O-O fragment. The ν(Fe-O) mode is observed at 559 cm-1 (H2O). Furthermore, both modes shift down by 3 cm-1, documenting the formulation as a hydroperoxo complex, in agreement with EPR data.

AB - The resonance Raman spectra of the hydroperoxo complex of camphor-bound CYP101 have been obtained by cryoradiolytic reduction of the oxygenated ferrous form that had been rapidly frozen in water/glycerol frozen solution; EPR spectroscopy was employed to confirm the identity of the trapped intermediate. The ν(O-O) mode, appearing at 799 cm-1, is observed for the first time in a peroxo-heme adduct. It is assigned unambiguously by employing isotopomeric mixtures of oxygen gas containing 50% 16O18O, confirming the presence of an intact O-O fragment. The ν(Fe-O) mode is observed at 559 cm-1 (H2O). Furthermore, both modes shift down by 3 cm-1, documenting the formulation as a hydroperoxo complex, in agreement with EPR data.

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JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

IS - 20

ER -

Resonance Raman detection of the hydroperoxo intermediate in the cytochrome P450 enzymatic cycle

Abstract

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