Hybrid In Silico/In Vitro Approaches for the Identification of Functional Cholesterol-Binding Domains in Membrane Proteins
Hybrid In Silico/In Vitro Approaches for the Identification of Functional Cholesterol-Binding Domains in Membrane Proteins
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DOI：
10.1007/978-1-4939-6875-6_2
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发表时间：
2017-01-01
期刊：
影响因子：
--
通讯作者：
Fantini, Jacques
中科院分区：
文献类型：
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作者：
Di Scala, Coralie;Fantini, Jacques
在真核细胞中，胆固醇是多种膜蛋白的重要调节剂，包括受体、转运体和离子通道。了解胆固醇如何与膜蛋白相互作用是一项艰巨的任务，因为这些蛋白质与胆固醇复合物的结构数据很少。在这里，我们描述了一种基于蛋白质-胆固醇相互作用的计算机研究的双重方法，结合了蛋白质插入含胆固醇单层的物理化学测量。通过仔细分析预测的胆固醇结合位点内外的关键突变的影响，我们的算法得到了验证。我们的方法是通过对胆固醇与阿尔茨海默氏症β -淀粉样肽结合的完整分析来说明的，β -淀粉样肽是一种通过胆固醇依赖过程穿透脑细胞质膜的蛋白质。
In eukaryotic cells, cholesterol is an important regulator of a broad range of membrane proteins, including receptors, transporters, and ion channels. Understanding how cholesterol interacts with membrane proteins is a difficult task because structural data of these proteins complexed with cholesterol are scarce. Here, we describe a dual approach based on in silico studies of protein-cholesterol interactions, combined with physico-chemical measurements of protein insertion into cholesterol-containing monolayers. Our algorithm is validated through careful analysis of the effect of key mutations within and outside the predicted cholesterol-binding site. Our method is illustrated by a complete analysis of cholesterol-binding to Alzheimer's beta-amyloid peptide, a protein that penetrates the plasma membrane of brain cells through a cholesterol-dependent process.