Josic Djuro, Brown Mari Kino, Huang Feilei, Callanan Helen, Rucević Marijana, Nicoletti Alison, Clifton James, Hixson Douglas C
Proteomics Core, COBRE Center for Cancer Research Development, Rhode Island Hospital, The CORO Center, Providence, RI 02903, USA.
Electrophoresis. 2005 Jul;26(14):2809-22. doi: 10.1002/elps.200500060.
A model system for selective solubilization and fast separation of proteins from the rat liver membrane fraction and purified rat liver plasma membranes for their further proteomic analysis is presented. For selective solubilization, high-pH solutions and a concentrated urea solution, combined with different detergents, are used. After extraction, proteins are separated by anion-exchange chromatography or a combination of anion- and cation-exchange chromatography with convective interaction monolithic supports. This separation method enables fast and effective prefractionation of membrane proteins based on their hydrophobicity and charge prior to one-dimensional (1-D) and 2-D electrophoresis and mass spectrometry. By use of this sample preparation method, the less-abundant proteins can be detected and identified.
本文介绍了一种用于从大鼠肝膜组分和纯化的大鼠肝质膜中选择性溶解和快速分离蛋白质以进行进一步蛋白质组学分析的模型系统。对于选择性溶解，使用高pH溶液、浓缩尿素溶液以及不同的去污剂。提取后，通过阴离子交换色谱或阴离子和阳离子交换色谱与对流相互作用整体柱的组合来分离蛋白质。这种分离方法能够在一维（1-D）和二维电泳以及质谱分析之前，基于膜蛋白的疏水性和电荷对其进行快速有效的预分级分离。通过使用这种样品制备方法，可以检测和鉴定低丰度蛋白质。
