Reconstitution of membrane proteins: catalysis by cholesterol of insertion of integral membrane proteins into preformed lipid bilayers.
The presence of cholesterol in small unilamellar vesicles (ULV) of dimyristoylphosphatidylcholine (DMPC) catalyzes fusion of the vesicles at temperatures below the upper limit for the gel to liquid-crystalline phase transition of the DMPC. The extent to which ULV grow depends on the concentration of cholesterol in the vesicles and on temperature. Maximum growth occurs at 21 degrees C. It decreases as the temperature is lowered below 21 degrees C. Growth does not occur at temperatures above the phase transition. In addition, the presence of cholesterol in ULV of DMPC catalyzes the insertion of integral membrane proteins into the vesicles. Thus, bacteriorhodopsin from Halobacterium halobrium, UDPglucuronosyltransferase (EC 2.4.1.17) from pig liver microsomes, and cytochrome oxidase from beef heart mitochondria formed stable lipid-protein complexes spontaneously when added to ULV containing cholesterol at temperatures under which these vesicles would fuse. Incorporation of these proteins into the ULV of DMPC did not occur in the absence of cholesterol or in the presence of cholesterol when the temperature of the system was above that for the phase transition. It appears that cholesterol lowers the energy barrier for fusion of ULV of DMPC and for insertion of integral membrane proteins into these bilayers. Studies with bacteriorhodopsin suggest that the energy barrier for insertion of proteins into ULV containing cholesterol is smaller than the energy barrier for fusion of the ULV with each other.
D000818:Animals; D001436:Bacteriorhodopsins / Q000378:metabolism; D002417:Cattle; D002784:Cholesterol / Q000378:metabolism*; D004134:Dimyristoylphosphatidylcholine; D003576:Electron Transport Complex IV / Q000378:metabolism; D014453:Glucuronosyltransferase / Q000378:metabolism; D006217:Halobacterium / Q000378:metabolism; D007700:Kinetics; D008051:Lipid Bilayers; D008565:Membrane Proteins / Q000378:metabolism*; D008854:Microscopy, Electron; D008862:Microsomes, Liver / Q000201:enzymology; D008929:Mitochondria, Heart / Q000201:enzymology; D013552:Swine; D013816:Thermodynamics
D008051:Lipid Bilayers; D008565:Membrane Proteins; D001436:Bacteriorhodopsins; D002784:Cholesterol; D003576:Electron Transport Complex IV; D014453:Glucuronosyltransferase; D004134:Dimyristoylphosphatidylcholine