PMID: 8611604Jan 12, 1996Paper

Effect of cholesterol on the tight insertion of cytochrome b5 into large unilamellar vesicles

Biochimica Et Biophysica Acta
K M Taylor, M A Roseman

Abstract

When cytochrome b5 is added to large unilamellar vesicles (LUVs) of 1-palmitoyl-2-oleoylphosphatidylcholine (POPC), it binds predominantly in a 'loose,' or transferable form. Prolonged incubation of 30 degrees C leads to insertion in the physiological 'tight,' nontransferable form, with a halftime for the loose --> tight conversion of approx. 9 days. In this study, the effect of cholesterol on the rate of tight insertion was determined. Tight binding was assayed by depleting the LUVs of loose cytochrome b5 with an excess of SUV acceptors and then separating the liposome populations by gel-filtration or velocity sedimentation. Incorporation of cholesterol into the LUVs was found to markedly increase the rate of tight insertion, even though cholesterol decreases the equilibrium binding constant and saturation level of protein binding. The effect is not a continuously increasing function of cholesterol content, but attains a maximum at 20-25% mol%, where the rate enhancement is approx. 10-fold over baseline. At higher cholesterol levels, the rate decreases, returning to baseline at 40 mol% cholesterol. These observations are highly unusual in that cholesterol generally decreases the membrane binding affinity and the permeability o...Continue Reading

References

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Citations

Feb 15, 2003·Journal of Pharmaceutical Sciences·Nikhil O Dhoot, Margaret A Wheatley
Sep 2, 2008·Langmuir : the ACS Journal of Surfaces and Colloids·Huong Thi BuiRyoichi Kuboi

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