Abstract:
Numerous bioinformatics tools allow predicting the localization of membrane proteins in the outer or inner membrane of Escherichia coli with high precision. Nevertheless, it might be desirable to experimentally verify such predictions or to assay the correct localization of recombinant or mutated variants of membrane proteins. Here we describe two methods (preferential detergent solubilization and sucrose-gradient fractionation) that allow to fractionate Gram-negative bacterial membranes and subsequently to enrich inner or outer membrane proteins.
摘要:
许多生物信息学工具可以高精度地预测膜蛋白在大肠杆菌外膜或内膜中的定位。然而,可能需要通过实验验证此类预测或测定膜蛋白的重组或突变变体的正确定位。在这里,我们描述了两种方法(优先去污剂溶解和蔗糖梯度分馏),允许分馏革兰氏阴性细菌膜并随后富集内或外膜蛋白。