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Biophys. J. BioFAST: First Published February 18, 2005. doi:10.1529/biophysj.104.051870
© 2005 by the Biophysical Society.

A more recent version of this article appeared on May 1, 2005.
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MEMBRANES

Structure of cholesterol/ceramide monolayer mixtures. Implications to the molecular organization of lipid rafts

Luana Scheffer 1, Inna Solomonov 1, Markus Jan Weygand 2, Kristian Kjaer 3, Leslie Leiserowitz 1 and Lia Addadi 1*

1 Weizmann Institute of Science
2 Max Planck Institute of Colloids & Interfaces
3 Risø National Laboratory

* To whom correspondence should be addressed. E-mail: lia.addadi{at}weizmann.ac.il.

Submitted on August 25, 2004
Revised on September 27, 2004
Accepted on 26 January 2005


  Abstract
The structure of monolayers of cholesterol/ceramide mixtures was investigated using grazing incidence X-ray diffraction (GIXD), immuno-fluorescence and atomic force microscopy (AFM) techniques. GIXD measurements showed the existence of a crystalline mixed phase of the two components within a range of compositions of cholesterol:ceramide between 100:0 and 67:33. The mixed phase coexists with a ceramide crystalline phase in the range of compositions between 50:50 and 30:70; between 30:70 and 0:100 only a crystalline phase of ceramide was detected. The latter was determined and modeled. Immuno-labeling was performed with an antibody specific to the cholesterol monohydrate crystalline arrangement. The antibody recognizes crystalline cholesterol monolayers, but does not interact with crystalline ceramide. Immuno-fluorescence and AFM data show that in uncompressed ceramide monolayers, the highly crystalline phase coexists with a disordered loosely packed phase. In contrast, no disordered phase coexists with the new crystalline mixed phase. We conclude that the new mixed phase represents a stable homogeneous arrangement of cholesterol with ceramide. As ceramide incorporates the lipid backbone common to all sphingolipids, this arrangement may be relevant to the understanding of the molecular organization of lipid rafts.

Key Words: antibodies, cholesterol rafts, cholesterol-rich domains, grazing incidence diffractions, immunofluorescence, membrane organization




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Copyright © 2005 by the Biophysical Society.