The influence of cholesterol incorporation and removal on lipid-bilayer viscosity and electron transfer in rat-liver microsomes.
The incorporation and removal of cholesterol from rat liver microsomes was used as a methodical approach to investigate the molecular organization of microsomal redox-chains. It was shown that the incorporation of cholesterol in microsomes increases and removal of cholesterol decreases lipid bilayer viscosity as indicated from the rate of fluorescent probe-pyrene eximerisation in cholesterol-enriched and cholesterol-depleted microsomes. The increase of membrane viscosity slows down the initial rates and decreases the rate constants of cytochrome b5 reduction by NAD(P)H, whereas the decrease of membrane viscosity enhances the initial rates and increases the rate constants of these reactions. The rates of cytochrome P450 reduction by reduced pyridine nucleotides do not depend on the viscosity of lipid bilayer. The incorporation and removal of cholesterol from microsomes was not followed by any essential changes in the rates of dimethylaniline N-demethylation, aniline p-hydroxylation, p-nitroanisole O-demethylation, oxygen consumption, oxidation of NADH and NADPH. Thus the reduction of cytochrome b5 by NADH and NADPH is the diffusion-dependent reaction in the redox-chains of microsomes only.
D000818:Animals; D002784:Cholesterol / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D003573:Cytochrome b Group / Q000378:metabolism; D015786:Cytochromes b5; D004579:Electron Transport; D066298:In Vitro Techniques; D007425:Intracellular Membranes / Q000378:metabolism; D008051:Lipid Bilayers / Q000378:metabolism*; D008297:Male; D008560:Membrane Fluidity; D008862:Microsomes, Liver / Q000201:enzymology / Q000378:metabolism*; D009243:NAD / Q000378:metabolism; D010084:Oxidation-Reduction; D010101:Oxygen Consumption; D051381:Rats; D013050:Spectrometry, Fluorescence; D014783:Viscosity
D003573:Cytochrome b Group; D008051:Lipid Bilayers; D009243:NAD; D015786:Cytochromes b5; D003577:Cytochrome P-450 Enzyme System; D002784:Cholesterol