Leyton Denisse L, Belousoff Matthew J, Lithgow Trevor
Research School of Biology, Australian National University, Canberra, ACT, 0200, Australia.
Department of Microbiology, Monash University, Building 77, 23 Innovation Walk, Clayton Campus, Melbourne, VIC, 3800, Australia.
Methods Mol Biol. 2015;1329:1-16. doi: 10.1007/978-1-4939-2871-2_1.
The outer membranes of gram-negative bacteria contain integral membrane proteins, most of which are of β-barrel structure, and critical for bacterial survival. These β-barrel proteins rely on the β-barrel assembly machinery (BAM) complex for their integration into the outer membrane as folded species. The central and essential subunit of the BAM complex, BamA, is a β-barrel protein conserved in all gram-negative bacteria and also found in eukaryotic organelles derived from bacterial endosymbionts. In Escherichia coli, BamA docks with four peripheral lipoproteins, BamB, BamC, BamD and BamE, partner subunits that add to the function of the BAM complex in outer membrane protein biogenesis. By way of introduction to this volume, we provide an overview of the work that has illuminated the mechanism by which the BAM complex drives β-barrel assembly. The protocols and methodologies associated with these studies as well as the challenges encountered and their elegant solutions are discussed in subsequent chapters.
革兰氏阴性菌的外膜含有整合膜蛋白,其中大多数是β桶状结构,对细菌的存活至关重要。这些β桶状蛋白依赖β桶组装机器(BAM)复合体以折叠形式整合到外膜中。BAM复合体的核心且必需的亚基BamA是一种β桶状蛋白,在所有革兰氏阴性菌中都保守,在源自细菌内共生体的真核细胞器中也有发现。在大肠杆菌中,BamA与四种外周脂蛋白BamB、BamC、BamD和BamE对接,这些伙伴亚基增强了BAM复合体在外膜蛋白生物合成中的功能。作为对本卷的介绍,我们概述了阐明BAM复合体驱动β桶组装机制的研究工作。后续章节将讨论与这些研究相关的实验方案和方法,以及遇到的挑战和巧妙的解决方案。