Choul-Li Souhaila, Adams Hendrik, Pattus Franc, Celia Hervé
Département Récepteurs et Protéines Membranaires, UMR7175-LC1, ESBS, Illkirch, France.
Electrophoresis. 2008 Mar;29(6):1333-8. doi: 10.1002/elps.200700612.
Horizontal nondenaturing electrophoresis of proteins in polyacrylamide gels was used to observe specific interactions between membrane proteins. The method was particularly well suited for solubilized transporters of the outer membrane of Gram-negative bacteria, and allowed specific complexes of transporter and the inner-membrane protein TonB to be isolated. We have used this method to investigate the interactions between four different outer-membrane transporters, and the TonB proteins from two different organisms. The results show that a stable complex can be isolated on gels for all the proteins studied, but can depend in some cases of the detergent used for solubilization. Furthermore, we observe cross-species interaction as TonB from a given organism can interact with transporters from another organism.
采用聚丙烯酰胺凝胶中蛋白质的水平非变性电泳来观察膜蛋白之间的特异性相互作用。该方法特别适用于革兰氏阴性菌外膜中溶解的转运蛋白，并能分离出转运蛋白与内膜蛋白托普辛（TonB）的特异性复合物。我们已使用该方法研究了四种不同外膜转运蛋白与来自两种不同生物体的托普辛蛋白之间的相互作用。结果表明，对于所有研究的蛋白质，都能在凝胶上分离出稳定的复合物，但在某些情况下可能取决于用于溶解的去污剂。此外，我们观察到跨物种相互作用，因为来自给定生物体的托普辛可以与来自另一种生物体的转运蛋白相互作用。
