Hooker Brian S, Bigelow Diana J, Lin Chiann-Tso
Pacific Northwest National Laboratory, Richland, Washington, USA.
Biochem Biophys Res Commun. 2007 Nov 23;363(3):457-61. doi: 10.1016/j.bbrc.2007.09.031. Epub 2007 Sep 19.
Nearly one-third of all genes in various organisms encode membrane-associated proteins that participate in numerous protein-protein interactions important to the processes of life. However, membrane protein interactions pose significant challenges due to the need to solubilize membranes without disrupting protein-protein interactions. Traditionally, analysis of isolated protein complexes by high-resolution 2D gel electrophoresis has been the main method used to obtain an overall picture of proteome constituents and interactions. However, this method is time consuming, labor intensive, detects only abundant proteins and is limited with respect to the coverage required to elucidate large interaction networks. In this review, we discuss the application of various methods to elucidate interactions involving membrane proteins. These techniques include methods for the direct isolation of single complexes or interactors as well as methods for characterization of entire subcellular and cellular interactomes.
各种生物体中近三分之一的基因编码与膜相关的蛋白质，这些蛋白质参与了许多对生命过程至关重要的蛋白质-蛋白质相互作用。然而，由于需要在不破坏蛋白质-蛋白质相互作用的情况下溶解膜，膜蛋白相互作用带来了重大挑战。传统上，通过高分辨率二维凝胶电泳分析分离的蛋白质复合物一直是用于全面了解蛋白质组组成和相互作用的主要方法。然而，这种方法耗时、费力，只能检测到丰富的蛋白质，并且在阐明大型相互作用网络所需的覆盖范围方面存在局限性。在这篇综述中，我们讨论了各种方法在阐明涉及膜蛋白的相互作用中的应用。这些技术包括直接分离单个复合物或相互作用分子的方法，以及表征整个亚细胞和细胞相互作用组的方法。
