Institute of Molecular Pathology, Faculty of Military Health Sciences, University of Defense, Trebesska 1575, 500 01 Hradec Kralove, Czech Republic.
J Proteomics. 2011 Dec 10;75(1):257-69. doi: 10.1016/j.jprot.2011.05.006. Epub 2011 May 11.
The study of membrane proteins and membrane protein complexes (MPC) provides crucial information in the field of bacterial physiology and pathogenesis research. The method of blue native polyacrylamide gel electrophoresis and its combination with SDS-PAGE (BN/SDS-PAGE) were here employed to study the membrane complexome of an intracellular bacterium Francisella tularensis, the causative agent of a severe disease tularemia. In the presented study we describe the subunit composition and stoichiometry of several MPC involved in various cell functions (oxidative phosphorylation, membrane transport, cell division, membrane or periplasmic proteins folding, iron storage, phospholipid and cell envelope biosynthesis). Moreover, some undocumented or hypothetical MPC with possible connection to virulence factors were also proposed and some newly detected subunits were assigned to known complexes. The BN/SDS-PAGE combined with mass spectrometry appeared to be a strong tool in the investigation of membrane proteins and complexes and thus successfully complements the traditional electrophoresis approaches.
研究膜蛋白和膜蛋白复合物（MPC）为细菌生理学和发病机制研究领域提供了关键信息。本研究采用蓝色非变性聚丙烯酰胺凝胶电泳及其与 SDS-PAGE 的结合（BN/SDS-PAGE）方法，研究了细胞内细菌土拉弗朗西斯菌（引起严重疾病土拉菌病的病原体）的膜复合物组。在本研究中，我们描述了几种参与各种细胞功能（氧化磷酸化、膜转运、细胞分裂、膜或周质蛋白折叠、铁储存、磷脂和细胞壁生物合成）的 MPC 的亚基组成和化学计量。此外，还提出了一些与毒力因子可能有关的未记录或假设的 MPC，并将一些新检测到的亚基分配到已知的复合物中。BN/SDS-PAGE 与质谱相结合似乎是研究膜蛋白和复合物的强大工具，因此成功地补充了传统的电泳方法。
