Mechanism of cholesterol and phosphatidylcholine exchange or transfer between unilamellar vesicles

• Published in: Biochemistry (Easton) 1981-05, Vol.20 (10), p.2893-2900
• Main Authors: McLean, L. R, Phillips, M. C
• Format: Article
• Language: English
• Subjects: Carbon Radioisotopes; Cholesterol; Dialysis; Egg Yolk; Female; Kinetics; Lipid Bilayers; Liposomes; Microscopy, Electron; Molecular Conformation; Phosphatidylcholines; Tritium
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi00513a028

The mechanism of cholesterol and phosphatidylcholine exchange has been investigated by following the transfer of radiolabeled cholesterol and phosphatidylcholine from negatively charged, unilamellar cholesterol-egg yolk phosphatidylcholine donor vesicles to neutral acceptor vesicles of similar composition. Vesicles were incubated in the absence of protein and were stable to fusion over the course of the experiment. At intervals, donor and acceptor vesicles were separated by passage through a column of DEAE-Sepharose; less than 1% of the charged and 80-95% of the neutral vesicles were recovered in the eluate. Over 12 h at 37 degrees C, 90% of the donor vesicle [4-14C]cholesterol was transferred to the acceptor vesicles in a first-order process whose half-time was 2.3 +/- 0.3 h. This indicates that transfer of cholesterol molecules from the inner to outer monolayer of the vesicle bilayer is not rate limiting in exchange. In contrast to cholesterol exchange, the half-time for 1-palmitoyl-2-oleoyl[1-14C]phosphatidylcholine exchange was 48 +/- 5 h so that more than six molecules of cholesterol were transferred for each molecule of phosphatidylcholine. The interfacial flux of cholesterol from the donor bilayer is 5.3 x 10(-15) mol cm-2 s-1 (approximately 3 molecules/min for an average vesicle) and is similar to fluxes observed in other systems where phosphatidylcholine or cholesterol ester exchange is catalyzed by an exchange protein. When the acceptor vesicle concentration was increased 20-fold in cholesterol exchange experiments or 9-fold in phosphatidylcholine exchange experiments, the rate of label transfer was not affected. The activation energy of cholesterol exchange between 15 and 37 degrees C was 73 +/- 5 kJ mol-1. Transfer of cholesterol across a dialysis membrane is shown to be a slow process whose rate may be predicted by application of Fick's first law of diffusion. These results are only consistent with a mechanism of lipid exchange in which cholesterol and phosphatidylcholine diffuse through the aqueous phase; the experimental activation energy is associated with desorption of lipid from the donor bilayer into the aqueous phase.