AUTHOR=Schleicher Lena , Muras Valentin , Claussen Björn , Pfannstiel Jens , Blombach Bastian , Dibrov Pavel , Fritz Günter , Steuber Julia 

TITLE=Vibrio natriegens as Host for Expression of Multisubunit Membrane Protein Complexes

JOURNAL=Frontiers in Microbiology

VOLUME=9

YEAR=2018

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2018.02537

DOI=10.3389/fmicb.2018.02537

ISSN=1664-302X

ABSTRACT=<p><italic>Escherichia coli</italic> is a convenient host for the expression of proteins, but the heterologous production of large membrane protein complexes often is hampered by the lack of specific accessory genes required for membrane insertion or cofactor assembly. In this study we introduce the non-pathogenic and fast-growing <italic>Vibrio natriegens</italic> as a suitable expression host for membrane-bound proteins from <italic>Vibrio cholerae</italic>. We achieved production of the primary Na<sup>+</sup> pump, the NADH:quinone oxidoreductase (NQR), from <italic>V. cholerae</italic> in an active state, as indicated by increased overall NADH:quinone oxidoreduction activity of membranes from the transformed <italic>V. natriegens</italic>, and the sensitivity toward Ag<sup>+</sup>, a specific inhibitor of the NQR. Complete assembly of <italic>V. cholerae</italic> NQR expressed in <italic>V. natriegens</italic> was demonstrated by BN PAGE followed by activity staining. The secondary transport system Mrp from <italic>V. cholerae</italic>, another membrane-bound multisubunit complex, was also produced in <italic>V. natriegens</italic> in a functional state, as demonstrated by <italic>in vivo</italic> Li<sup>+</sup> transport. <italic>V. natriegens</italic> is a promising expression host for the production of membrane protein complexes from Gram-negative pathogens.</p>