Eubel Holger, Braun Hans-Peter, Millar A Harvey
ARC Centre of Excellence in Plant Energy Biology, University of Western Australia, 35 Stirling Hwy, Crawley 6009, Perth, Australia.
Plant Methods. 2005 Nov 16;1(1):11. doi: 10.1186/1746-4811-1-11.
Intact protein complexes can be separated by apparent molecular mass using a standard polyacrylamide gel electrophoresis system combining mild detergents and the dye Coomassie Blue. Referring to the blue coloured gel and the gentle method of solubilization yielding native and enzymatically active protein complexes, this technique has been named Blue-Native Polyacrylamide Gel-Electrophoresis (BN-PAGE). BN-PAGE has become the method of choice for the investigation of the respiratory protein complexes of the electron transfer chains of a range of organisms, including bacteria, yeasts, animals and plants. It allows the separation in two dimensions of extremely hydrophobic protein sets for analysis and also provides information on their native interactions. In this review we discuss the capabilities of BN-PAGE in proteomics and the wider investigation of protein:protein interactions with a focus on its use and potential in plant science.
完整的蛋白质复合物可以通过使用结合了温和去污剂和考马斯亮蓝染料的标准聚丙烯酰胺凝胶电泳系统,根据表观分子量进行分离。鉴于蓝色凝胶以及温和的溶解方法能产生天然且具有酶活性的蛋白质复合物,这项技术被命名为蓝色天然聚丙烯酰胺凝胶电泳(BN-PAGE)。BN-PAGE已成为研究包括细菌、酵母、动物和植物在内的一系列生物体电子传递链呼吸蛋白复合物的首选方法。它能够对极其疏水的蛋白质组进行二维分离以用于分析,还能提供有关其天然相互作用的信息。在本综述中,我们将讨论BN-PAGE在蛋白质组学方面的能力以及对蛋白质:蛋白质相互作用的更广泛研究,重点关注其在植物科学中的应用和潜力。