Rotation of cytochrome P-450. II. Specific interactions of cytochrome P-450 with NADPH-cytochrome P-450 reductase in phospholipid vesicles.
作者： J Gut , C Richter , R J Cherry , K H Winterhalter , S Kawato
DOI: 10.1016/S0021-9258(18)34533-2
关键词:
摘要: Purified rat liver microsomal cytochrome P-450 and NADPH-cytochrome reductase were co-reconstituted in phosphatidylcholine-phosphatidylethanolamine-phosphatidylserine vesicles using a cholate dialysis technique. The co-reconstitution of the enzymes was demonstrated proteoliposomes fractionated by centrifugation glycerol gradient. catalyzed N-demethylation variety substrates. Rotational diffusion measured detecting decay absorption anisotropy r(t), after photolysis heme.CO complex vertically polarized laser flash. rotational mobility P-450, when reconstituted alone, found to be dependent on lipid protein ratio weight (L/P450) (Kawato, S., Gut, J., Cherry, R. Winterhalter, K. H., Richter, C. (1982) J. Biol. Chem. 257, 7023-7029). About 35% immobilized rest rotating with mean relaxation time phi 1 about 95 mus L/P450 = vesicle. In 10 vesicles, 10% immobile congruent 55 mus. Co-reconstitution equimolar amounts into above results completely mobile 40 Only small decrease fraction is observed molar 5. suggest formation monomolecular 1:1 between liposomes.
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