Rabilloud Thierry
Laboratoire, d'immunologie, INSERM, Grenoble, France.
Methods Mol Biol. 2009;528:259-67. doi: 10.1007/978-1-60327-310-7_18.
Because of the outstanding ability of two-dimensional electrophoresis to separate complex mixtures of intact proteins, it would be advantageous to apply it to all types of proteins, including hydrophobic and membrane proteins. Unfortunately, poor solubility hampers the analysis of these molecules. As these problems arise mainly in the extraction and isoelectric focusing steps, the solution is to improve protein solubility under the conditions prevailing during isoelectric focusing. This chapter describes the use of chaotropes and novel detergents to enhance protein solubility during sample extraction and isoelectric focussing, and discusses the contribution of these compounds to improving proteomic analysis of membrane proteins.
由于二维电泳在分离完整蛋白质复杂混合物方面具有卓越能力,将其应用于所有类型的蛋白质(包括疏水蛋白和膜蛋白)会很有利。不幸的是,溶解性差阻碍了对这些分子的分析。由于这些问题主要出现在提取和等电聚焦步骤中,解决办法是在等电聚焦过程中普遍存在的条件下提高蛋白质的溶解度。本章描述了在样品提取和等电聚焦过程中使用离液剂和新型去污剂来提高蛋白质溶解度,并讨论了这些化合物对改善膜蛋白蛋白质组学分析的作用。