Cyclodextrin-catalyzed extraction of fluorescent sterols from monolayer membranes and small unilamellar vesicles.
This study examined the kinetics of sterol desorption from monolayer and small unilamellar vesicle membranes to 2-hydroxypropyl-beta-cyclodextrin. The sterols used include cholesterol, dehydroergosterol (ergosta-5,7,9,(11),22-tetraen-3beta-ol) and cholestatrienol (cholesta-5,7,9,(11)-trien-3beta-ol). Desorption rates of dehydroergosterol and cholestatrienol from pure sterol monolayers were faster (3.3-4.6-fold) than the rate measured for cholesterol. In mixed monolayers (sterol: 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine 30:70 mol%), both dehydroergosterol and cholestatrienol desorbed faster than cholesterol. clearly indicating a difference in interfacial behavior of these sterols. In vesicle membranes desorption of dehydroergosterol was slower than desorption of cholestatrienol, and both rates were markedly affected by the phospholipid composition. Desorption of sterols was slower from sphingomyelin as compared to phosphatidylcholine vesicles. Desorption of fluorescent sterols was also faster from vesicles prepared by ethanol-injection as compared to extruded vesicles. The results of this study suggest that dehydroergosterol and cholestatrienol differ from cholesterol in their membrane behavior, therefore care should be exercised when experimental data derived with these probes are interpreted.
Ohvo-Rekilä
H
H
epartment of Biochemistry and Pharmacy, Abo Akademi University, Turku, Finland. hohvo@abo.fi
Akerlund
B
B
Slotte
J P
JP
D000073738:2-Hydroxypropyl-beta-cyclodextrin; D002782:Cholestenes / Q000737:chemistry; D002784:Cholesterol / Q000737:chemistry; D003505:Cyclodextrins / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004875:Ergosterol / Q000031:analogs & derivatives / Q000737:chemistry; D000431:Ethanol / Q000494:pharmacology; D007700:Kinetics; D008567:Membranes, Artificial; D010713:Phosphatidylcholines / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013261:Sterols / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism*; D013997:Time Factors; D014867:Water / Q000378:metabolism; D047392:beta-Cyclodextrins
D002782:Cholestenes; D003505:Cyclodextrins; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D013261:Sterols; D047392:beta-Cyclodextrins; D014867:Water; C010905:dehydroergosterol; D000073738:2-Hydroxypropyl-beta-cyclodextrin; D000431:Ethanol; C025498:cholesta-5,7,9-trien-3 beta-ol; D002784:Cholesterol; C028694:1-palmitoyl-2-oleoylphosphatidylcholine; D004875:Ergosterol