Strauss A, Pohlner J, Klauser T, Meyer T F
Max-Planck-Institut für Biologie, Abteilung Infektionsbiologie, Tübingen, Germany.
FEMS Microbiol Lett. 1995 Apr 1;127(3):249-54. doi: 10.1111/j.1574-6968.1995.tb07481.x.
A glycine-histidine tag (Gly3His6) was added to the C-terminus of a fusion protein consisting of the cholera toxin B-subunit (CtxB) and the IgA protease beta-domain (Iga beta). The aim was to facilitate single-step purification and to create a suitable tool for kinetic and structural studies on Iga beta-driven protein translocation across the outer membrane of Gram-negative bacteria. We demonstrate that the glycine-histidine tag does not interfere with the assembly of Iga beta in the outer membrane and that the translocator function of the modified Iga beta is maintained. The applicability of the new construct for the dissection of the Iga beta mediated translocation process and general aspects of C-terminal histidine tagging of outer membrane proteins are discussed.
在由霍乱毒素B亚基（CtxB）和IgA蛋白酶β结构域（Igaβ）组成的融合蛋白的C末端添加了一个甘氨酸-组氨酸标签（Gly3His6）。目的是促进单步纯化，并创建一个适用于对Igaβ驱动的蛋白质跨革兰氏阴性菌外膜转运进行动力学和结构研究的工具。我们证明，甘氨酸-组氨酸标签不会干扰Igaβ在外膜中的组装，并且修饰后的Igaβ的转运功能得以维持。讨论了新构建体在剖析Igaβ介导的转运过程中的适用性以及外膜蛋白C末端组氨酸标签的一般情况。
