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C100654200v1
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Papers In Press, published online ahead of print November 20, 2001
J. Biol. Chem, 10.1074/jbc.C100654200
Submitted on November 13, 2001
Revised on November 20, 2001
Accepted on November 19, 2001

Cholesterol is not crucial for the existence of microdomains in kidney brush border membrane models

Pierre Emmanuel Milhiet, Marie-Cecile Giocondi, and Christian Le Grimellec

Centre de Biochimie Structurale, UMR554 INSERM, Montpellier 34090

Corresponding Author: pem{at}cbs.univ-montp1.fr

The external membrane leaflet plays a key role in the organization of the cell plasma membrane as a mosaic of ordered microdomains enriched in sphingolipids and cholesterol and of fluid domains. In this study, the thermotropic behavior and the topology of bilayers made of a phosphatidylcholine/sphingomyelin mixture, which mimicks the lipid composition of the external leaflet of renal brush border membranes, were examined by differential scanning calorimetry and atomic force microscopy. In the absence of cholesterol, a broad phase separation process occurred where ordered gel phase domains of size varying from the mesoscopic to the microscopic scale, enriched in sphingomyelin, occupied half of the bilayer surface at room temperature. Increasing amounts of cholesterol progressively decreased the enthalpy of the transition and modified the topology of membranes domains up to a concentration of 33 mol % for which no membrane domains were detected. These results strongly suggest that, in membranes highly enriched in sphingolipids like renal and intestinal brush borders, there is a threshold close to the physiological concentration above which cholesterol acts as a suppressor rather than as a promoter of membrane domains. They also suggest that cholesterol depletion does not abolish the lateral heterogenity in brush border membranes.


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