CHAPSTEROL - A novel cholesterol-based detergent
CHAPSTEROL - A novel cholesterol-based detergent
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DOI：
10.1111/j.1742-4658.2004.04517.x
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发表时间：
2005-02-01
期刊：
影响因子：
5.4
通讯作者：
Gimpl, G
中科院分区：
文献类型：
--
作者：
Gehrig-Burger, K;Kohout, L;Gimpl, G
介绍了一种新型胆固醇清洗剂CHAPSTEROL的设计、合成和表征，该清洗剂用于功能增溶胆固醇依赖的膜蛋白。为了验证CHAPSTEROL，我们使用了催产素受体，一种需要胆固醇作为其高亲和力结合状态的G蛋白偶联受体。使用可光激活的胆固醇类似物[H-3]6，6-偶氮胆烷-3-β-醇[3alphaH]，我们证明了CHAPSTEROL的增溶作用导致胆固醇结合蛋白的丰富，而广泛使用的胆汁酸衍生物CHAPSO则导致胆固醇结合蛋白的显著减少。与Triton X-100和CHAPS类似，CHAPSTEROL维持小窝蛋白以及胆固醇和鞘磷脂在脂筏上的定位，即质膜上不溶于洗涤剂的微区。这些数据表明，CHAPSTEROL是一种适合于增溶胆固醇依赖的膜蛋白和分离RAFT的洗涤剂。
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 [H-3]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.