Darwiche Rabih, Schneiter Roger
Division of Biochemistry, Department of Biology, University of Fribourg, Chemin du Musée 10, 1700, Fribourg, Switzerland.
Methods Mol Biol. 2017;1645:361-368. doi: 10.1007/978-1-4939-7183-1_25.
Sterols are major constituents of the plasma membrane of eukaryotic cells and serve as a precursor for several classes of signaling molecules, including steroids and hydroxy sterols. They maintain the functionality and permeability barrier of the plasma membrane through lipid-lipid and lipid-protein interactions. The S. cerevisiae pathogen-related yeast proteins 1, 2, and 3 (Pry) belong to a large protein superfamily known as CAP/SCP/TAPS. Members of this superfamily have been implicated in a wide variety of processes, including immune defense in mammals and plants, pathogen virulence, sperm maturation and fertilization, venom toxicity, and prostate and brain cancer. Pry proteins bind and export sterols in vivo and the purified Pry1 protein binds sterols and related small hydrophobic compounds in vitro. Here we describe a method to determine lipid binding of a purified protein in vitro.
甾醇是真核细胞质膜的主要成分，也是几类信号分子的前体，包括类固醇和羟基甾醇。它们通过脂质-脂质和脂质-蛋白质相互作用维持质膜的功能和渗透屏障。酿酒酵母病原体相关酵母蛋白1、2和3（Pry）属于一个名为CAP/SCP/TAPS的大型蛋白质超家族。该超家族的成员参与了多种过程，包括哺乳动物和植物的免疫防御、病原体毒力、精子成熟和受精、毒液毒性以及前列腺癌和脑癌。Pry蛋白在体内结合并输出甾醇，纯化的Pry1蛋白在体外结合甾醇和相关的小疏水化合物。在这里，我们描述了一种在体外测定纯化蛋白脂质结合的方法。
