CHAPSTEROL. A novel cholesterol-based detergent.
Design, synthesis and characterization of CHAPSTEROL, a novel cholesterol-based detergent developed for functional solubilization of cholesterol-dependent membrane proteins are described. To validate CHAPSTEROL, we employed the oxytocin receptor, a G protein-coupled receptor requiring cholesterol for its high-affinity binding state. Using the photoactivatable cholesterol analogue [3H]6,6-azocholestan-3beta-ol[3alphaH], we demonstrate that solubilization by CHAPSTEROL leads to an enrichment of cholesterol-binding proteins whereas the widely used bile acid derivative CHAPSO leads to a significant depletion of cholesterol-binding proteins. Similar to Triton X-100 and CHAPS, CHAPSTEROL maintains the localization of caveolin as well as cholesterol and sphingomyelin to lipid rafts, i.e. detergent-insoluble microdomains of the plasma membrane. The data suggest that CHAPSTEROL is an appropriate detergent for the solubilization of cholesterol-dependent membrane proteins and isolation of rafts.
Gehrig-Burger
Katja
K
Institute of Biochemistry, University of Mainz, Germany. kburger@uni-mainz.de
Kohout
Ladislav
L
Gimpl
Gerald
G
D002460:Cell Line; D002784:Cholesterol / Q000138:chemical synthesis* / Q000737:chemistry* / Q000378:metabolism; D003902:Detergents / Q000138:chemical synthesis* / Q000737:chemistry* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D019920:Photoaffinity Labels; D018045:Receptors, Oxytocin / Q000378:metabolism
D003902:Detergents; D019920:Photoaffinity Labels; D018045:Receptors, Oxytocin; C498014:chapsterol; D002784:Cholesterol