Rates of spontaneous exchange of synthetic radiolabeled sterols between lipid vesicles.
14C-labeled sterols with structural variation in the polar function [3 alpha-OH, 3-O(CH2)2O-(CH2)2O(CH2)2OH, 3 alpha-NH2, 3 beta-NH2, and 3-OC(O)CHN = N] and at the 7 position (7-oxo, 7 alpha-OH, and 7 beta-OH) were synthesized and incorporated into unilamellar vesicles for studies of the rates of transfer to an excess of acceptor vesicles. Cholesterol, cholestanol, and epicholesterol underwent full exchange in a single kinetic pool, and 90% of the 3 alpha-triethoxycholesterol was exchangeable in one pool. Biphasic kinetics with full exchangeability were observed for cholesterylamines, which bear a positive charge at the 3 position; the slow phase reflects the high activation energy for inner-to-outer leaflet movement of the charged lipid. Biphasic kinetics were also found for cholesteryl diazoacetate, indicating that this photoaffinity probe and cholesterol have different mechanisms of transfer. Sterols that are more hydrophilic than cholesterol as estimated by reversed-phase high-performance chromatography (elution with acetonitrile-2-propanol, 4:1 v/v, with varying proportions of water) gave faster exchange rates than cholesterol, whereas sterols that are more hydrophobic gave slower exchange rates. However, the rates of [14C]sterol desorption from the lipid-water interface are not correlated with the relative sterol hydrophobicity as estimated by the logarithm of the capacity factors using acetonitrile-2-propanol-water as the mobile phase. These studies suggest that the interaction of sterols with phospholipids provides the principal physical-chemical basis for determining the rates of spontaneous exchange of sterols between bilayers.
Kan
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Department of Chemistry and Biochemistry, Queens College, City University of New York, Flushing 11367.
Yan
J
J
Bittman
R
R
D015060:1,2-Dipalmitoylphosphatidylcholine / Q000378:metabolism; D002250:Carbon Radioisotopes; D004083:Cholestanol / Q000378:metabolism; D002784:Cholesterol / Q000031:analogs & derivatives* / Q000378:metabolism; D002788:Cholesterol Esters / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003979:Diazonium Compounds / Q000378:metabolism; D006888:Hydroxycholesterols / Q000378:metabolism; D007653:Ketocholesterols / Q000378:metabolism; D007700:Kinetics; D008081:Liposomes / Q000378:metabolism*; D008563:Membrane Lipids / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism*
D002250:Carbon Radioisotopes; D002788:Cholesterol Esters; D003979:Diazonium Compounds; D006888:Hydroxycholesterols; D007653:Ketocholesterols; D008081:Liposomes; D008563:Membrane Lipids; D010713:Phosphatidylcholines; D010743:Phospholipids; C052336:cholesteryl diazoacetate; C072855:cholesterylamine; D015060:1,2-Dipalmitoylphosphatidylcholine; D004083:Cholestanol; D002784:Cholesterol; C003001:7-ketocholesterol