Cholesterol-dependent insertion of glycosylphosphatidylinositol-anchored enzyme

Evidence is now accumulating that the plasma membrane is organized in different lipid and protein subdomains. Thus, glycosylphosphatidylinositol (GPI)-anchored proteins are proposed to be clustered in membrane microdomains enriched in cholesterol and sphingolipids, called rafts. By a detergent-media...

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Bibliographic Details
Published in:Biochimica et biophysica acta 2002-08, Vol.1564 (2), p.473-478
Main Authors: Morandat, Sandrine, Bortolato, Muriel, Roux, Bernard
Format: Article
Language:English
Subjects:
Alkaline phosphatase
Alkaline Phosphatase - chemistry
Animals
Cattle
Cell Membrane - chemistry
Cholesterol
Cholesterol - chemistry
Detergents
Glycosylphosphatidylinositol anchor
Glycosylphosphatidylinositols - chemistry
Liposome
Models, Chemical
Protein Conformation
Protein Structure, Tertiary
Proteolipids - chemistry
Reconstitution
Temperature
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Summary:Evidence is now accumulating that the plasma membrane is organized in different lipid and protein subdomains. Thus, glycosylphosphatidylinositol (GPI)-anchored proteins are proposed to be clustered in membrane microdomains enriched in cholesterol and sphingolipids, called rafts. By a detergent-mediated method, alkaline phosphatase, a GPI-anchored enzyme, was efficiently inserted into the membrane of sphingolipids- and cholesterol-rich liposomes as demonstrated by flotation in sucrose gradients. We have determined the enzyme extraluminal orientation. Using defined lipid components to assess the possible requirements for GPI-anchored protein insertion, we have demonstrated that insertion into membranes was cholesterol-dependent as the cholesterol addition increased the enzyme incorporation in simple phosphatidylcholine liposomes.
ISSN:0005-2736
0006-3002
1879-2642
DOI:10.1016/S0005-2736(02)00497-2