Schindler Jens, Nothwang Hans Gerd
Abteilung Tierphysiologie, Fachbereich Biologie, Technische Universität Kaiserslautern, Kaiserslautern, Germany.
Proteomics. 2006 Oct;6(20):5409-17. doi: 10.1002/pmic.200600243.
Plasma membranes (PMs) are of particular importance for all living cells. They form a selectively permeable barrier to the environment. Many essential tasks of PMs are carried out by their proteinaceous components, including molecular transport, cell-cell interactions, and signal transduction. Due to the key role of these proteins for cellular function, they take center-stage in basic and applied research. A major problem towards in-depth identification and characterization of PM proteins by modern proteomic approaches is their low abundance and immense heterogeneity in different cells. Highly selective and efficient purification protocols are hence essential to any PM proteome analysis. An effective tool for preparative isolation of PMs is partitioning in aqueous polymer two-phase systems. In two-phase systems, membranes are separated according to differences in surface properties rather than size and density. Despite their rare application to the fractionation of animal tissues and cells, they represent an attractive alternative to conventional fractionation protocols. Here, we review the principles of partitioning using aqueous polymer two-phase systems and compare aqueous polymer two-phase systems with other methods currently used for the isolation of PMs.
质膜（PMs）对所有活细胞都尤为重要。它们形成了一道对环境具有选择性渗透性的屏障。质膜的许多重要任务是由其蛋白质成分执行的，包括分子运输、细胞间相互作用和信号转导。由于这些蛋白质对细胞功能的关键作用，它们在基础研究和应用研究中占据核心地位。利用现代蛋白质组学方法深入鉴定和表征质膜蛋白质的一个主要问题是它们在不同细胞中的低丰度和巨大的异质性。因此，高度选择性和高效的纯化方案对于任何质膜蛋白质组分析都是必不可少的。用于质膜制备分离的一种有效工具是在水性聚合物双相系统中进行分配。在双相系统中，膜是根据表面性质的差异而不是大小和密度来分离的。尽管它们很少应用于动物组织和细胞的分级分离，但它们是传统分级分离方案的一种有吸引力的替代方法。在这里，我们综述了使用水性聚合物双相系统进行分配的原理，并将水性聚合物双相系统与目前用于质膜分离的其他方法进行比较。
