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Originally published as Biophys J. BioFAST on March 25, 2005.
doi:10.1529/biophysj.104.054718
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Biophysical Journal 88:4054-4063 (2005)
© 2005 The Biophysical Society

Domain Formation and Stability in Complex Lipid Bilayers as Reported by Cholestatrienol

Y. Jenny E. Björkqvist, Thomas K. M. Nyholm, J. Peter Slotte and Bodil Ramstedt

Department of Biochemistry and Pharmacy, Åbo Akademi University, Tykistökatu 6 A, FIN 20520 Turku, Finland

Correspondence: Address reprint requests to Bodil Ramstedt, Tel.: 358-2-2154816; Fax: 358-2-2154010; E-mail: boramste{at}abo.fi.

In this study, we used cholestatrienol (CTL) as a fluorescent reporter molecule to study sterol-rich Lo domains in complex lipid bilayers. CTL is a fluorescent cholesterol analog that mimics the behavior of cholesterol well. The ability of 12SLPC to quench the fluorescence of cholestatrienol gives a measure of the amount of sterol included in Lo domains in mixed lipid membranes. The stability of sterol-rich domains formed in complex lipid mixtures containing saturated sphingomyelins, phosphatidylcholines, or galactosylceramide as potential domain-forming lipids were studied. The amount of sterol associated with sterol-rich domains seemed to always increase with increasing temperature. The quenching efficiency was highly dependent on the domain-forming lipid present in complex lipid mixtures. Sphingomyelins formed stable sterol-enriched domains and were able to shield CTL from quenching better than the other lipids included in this study. The saturated phosphatidylcholines also formed sterol-rich domains, but the quenching efficiency in membranes with these was higher than with sphingomyelins and the domains melted at lower temperatures. PGalCer was not able to form sterol-enriched domains. However, we found that PGalCer stabilized sterol-rich domains formed in PSM-containing bilayers. Using a fluorescent ceramide analog, we also demonstrated that N-palmitoyl-ceramide displaced the sterol from sphingolipid-rich domains in mixed bilayer membranes.




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