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J. Biol. Chem., Vol. 266, Issue 16, 10246-10248, 06, 1991

Observation of the O-O stretching Raman band for cytochrome P-450cam under catalytic conditions

T Egawa, T Ogura, R Makino, Y Ishimura and T Kitagawa
Institute for Molecular Science, Okazaki National Research Institutes, Japan.

Dioxygen stretching (voo) Raman band was observed for the oxy form of Pseudomonas putida cytochrome P-450 (P-450cam) generated at room temperature under catalytic conditions, that is, in the presence of D- camphor, beta-NADH, putidaredoxin, and putidaredoxin reductase, by using the mixed flow transient Raman apparatus. At the same time the visible absorption spectra were monitored for the transient species. It was found that the voo frequency is little altered by binding of putidaredoxin to P-450cam, although the reduction rate of the oxy form becomes faster. Another intermediate with an oxygen isotope-sensitive band was not found in a time region until 2 s after mixing of the reduced enzyme with oxygen.
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