Cytochrome P-450 rotamers control mixed-function oxygenation in reconstituted membranes. Rotational diffusion studied by delayed fluorescence depolarizationd`

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Abstract
1. Rotational diffusion of purified rabbit-liver microsomal cytochrome P-450 LM2 in reconstituted lipid-vesicle membranes was investigated by measurement of timedependent polarized emission of delayed fluorescence. 2. Cytochrome P-450 labelled with diiodofluorescein iodoacetamide exhibited strict uniaxial rotation about the normal to the membrane. 3. Benzphetamine retards rotation, while reduction of the cytochrome P-450 substratecomplex accelerates rotation. 4. A model is proposed in which cytochrome P-450 forms disc-shaped rotamers immersed in the bilayer membrane to a depth which is varied by substrate-induced and redox state-dependent conformational changes. The model describes a regulating mechanism of the electron transfer-controlling mixed-function oxygenation.