Rotation and interaction with epoxide hydrase of cytochrome P-450 in proteoliposomes

作者: Y Ohta , H Sasabe , H U Etter , S Kawato , C Richter

DOI: 10.3929/ETHZ-A-000692255

关键词: PopulationCytochromeHemeproteinHemeEpoxide hydrolaseCytochrome P450PhosphatidylcholineStereochemistryEpoxideChemistry

摘要: Abstract Purified rat liver cytochrome P-450MC or P-450PB was co-reconstituted with epoxide hydrase in liposomal vesicles made of phosphatidylcholine, phosphatidylethanolamine, and phosphatidylserine at a lipid to protein weight ratio 5 by the cholate dialysis procedure. Rotational diffusion cytochromes measured observing decay absorption anisotropy, r(t), after photolysis heme.CO complex vertically polarized laser flash. Analysis r(t) based on "rotation-about-membrane-normal" model. The measurements were used investigate interactions hydrase. Different rotational mobilities two observed. amount mobile molecules 78% for 91% P-450PB, rest immobile within experimental time range 1 ms. In presence 85% 96% mobile. Cross-linking anti-epoxide antibodies resulted drastic immobilization cytochromes, reducing population 49% 60% P-450PB. relaxation times phi populations ranged from 210 283 microseconds. These results imply that both transiently associate membranes. Further analysis data showed angle between heme plane membrane is 48 degrees 62 degrees, different value 55 reported previously (Gut, J., Richter, C., Cherry, R. Winterhalter, K. H., Kawato, S. (1983) J. Biol. Chem. 258, 8588-8594).

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