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  • Title: Cyclodextrin-mediated removal of sterols from monolayers: effects of sterol structure and phospholipids on desorption rate.
    Author: Ohvo H, Slotte JP.
    Journal: Biochemistry; 1996 Jun 18; 35(24):8018-24. PubMed ID: 8672506.
    Abstract:
    In this study, we have examined a number of parameters which affect the rate of sterol desorption from a model membrane surface (a monolayer at the air/water interface) to cyclodextrins (CD) in the aqueous subphase. The desorption experiments were carried out at a constant lateral surface pressure with a zero-order trough, which allowed for a determination of desorption rates which were unaffected by monolayer substrate concentration. At a surface pressure of 20 mN/m (30 degrees C), 0.9 mM beta-CD caused a desorption of about 13 pmol of cholesterol per minute and square centimeter of monolayer area. The desorption of cholesterol proceeded linearly as a time function and was sensitive to the concentration of beta-CD in the subphase. The rate of cholesterol desorption increased as the monolayer surface pressure increased (3->35 mN/m) but decreased slightly with increasing temperature (15->30 degrees C). The rate of sterol desorption appeared to be influenced by the relative polarity of the sterols. Oxidized sterols desorbed significantly faster than cholesterol (e.g., 4-cholesten-3-one desorbed 8.4-fold faster than cholesterol), whereas less polar sterols desorbed at slower rates [e.g., 20(R)-isoheptyl-5-pregnen-3 beta-ol, a cholesterol analogue with a ten-carbon branched side chain, desorbed at 1/10 of the rate of cholesterol]. Cholesterol desorption from a monolayer membrane containing both cholesterol and a phospholipid was much slower than from a pure cholesterol monolayer. When the effect of dipalmitoylphosphatidylcholine and N-palmitoylsphingomyelin on cholesterol desorption rate was compared, it was found that cholesterol desorption was much more retarded from sphingomyelin monolayers as compared to that from phosphatidylcholine monolayers. Taken together, the results of this study show that the beta-CD-enhanced desorption of cholesterol (and other sterols) from monolayer membranes is influenced by the polarity of the desorbing molecules, as well as by lipid/lipid interactions in the membranes. Since beta-CD has no surface activity of its own, it appears to be a useful, nonintrusive catalyzer of cholesterol desorption and is expected to become a valuable probe in membrane and cell research.
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