Open AccessC‐terminal glycine‐histidine tagging of the outer membrane protein Iga β of Neisseria gonorrhoeae
Author(s) -
Strauss Andreas,
Pohlner Johannes,
Klauser Thomas,
Meyer Thomas F
Publication year - 1995
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1995.tb07481.x
Subject(s) - bacterial outer membrane , histidine , biology , biochemistry , neisseria gonorrhoeae , protein subunit , chemistry , microbiology and biotechnology , amino acid , escherichia coli , gene
A glycine‐histidine tag (Gly 3 His 6 ) was added to the C‐terminus of a fusion protein consisting of the cholera toxin B‐subunit (CtxB) and the IgA protease β‐domain (Iga β). The aim was to facilitate single‐step purification and to create a suitable tool for kinetic and structural studies on Iga β‐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 β in the outer membrane and that the translocator function of the modified Iga β is maintained. The applicability of the new construct for the dissection of the Iga β mediated translocation process and general aspects of C‐terminal histidine tagging of outer membrane proteins are discussed.