{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T17:49:18Z","timestamp":1791308958491,"version":"4.3.1"},"reference-count":55,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2013,4,9]],"date-time":"2013-04-09T00:00:00Z","timestamp":1365465600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Wellcome Trust"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecular Microbiology"],"published-print":{"date-parts":[[2013,5]]},"abstract":"<jats:title>Summary<\/jats:title><jats:p>The <jats:styled-content style=\"fixed-case\">MtrCDE<\/jats:styled-content> multidrug pump, from <jats:italic><jats:styled-content style=\"fixed-case\">N<\/jats:styled-content>eisseria gonorrhoeae<\/jats:italic>, is assembled from the inner and outer membrane proteins <jats:styled-content style=\"fixed-case\">MtrD<\/jats:styled-content> and <jats:styled-content style=\"fixed-case\">MtrE<\/jats:styled-content>, which are connected by the periplasmic membrane fusion protein <jats:styled-content style=\"fixed-case\">MtrC<\/jats:styled-content>. Although it is clear that <jats:styled-content style=\"fixed-case\">MtrD<\/jats:styled-content> delivers drugs to the channel of <jats:styled-content style=\"fixed-case\">MtrE<\/jats:styled-content>, it remains unclear how drug delivery and channel opening are connected. We used a vancomycin sensitivity assay to test for opening of the <jats:styled-content style=\"fixed-case\">MtrE<\/jats:styled-content> channel. Cells expressing <jats:styled-content style=\"fixed-case\">MtrE<\/jats:styled-content> or <jats:styled-content style=\"fixed-case\">MtrE<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">E<\/jats:styled-content>434<jats:styled-content style=\"fixed-case\">K<\/jats:styled-content> were insensitive to vancomycin; but became moderately and highly sensitive to vancomycin respectively, when coexpressed with <jats:styled-content style=\"fixed-case\">MtrC<\/jats:styled-content>, suggesting that the <jats:styled-content style=\"fixed-case\">MtrE<\/jats:styled-content> channel opening requires <jats:styled-content style=\"fixed-case\">MtrC<\/jats:styled-content> binding and is energy\u2010independent. Cells expressing wild\u2010type <jats:styled-content style=\"fixed-case\">MtrD<\/jats:styled-content>, in an <jats:styled-content style=\"fixed-case\">MtrCE<\/jats:styled-content> background, were vancomycin\u2010insensitive, but moderately sensitive in an <jats:styled-content style=\"fixed-case\">MtrCE<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">E<\/jats:styled-content>434<jats:styled-content style=\"fixed-case\">K<\/jats:styled-content> background. The mutation of residues involved in proton translocation inactivated <jats:styled-content style=\"fixed-case\">MtrD<\/jats:styled-content> and abolished drug efflux, rendered both <jats:styled-content style=\"fixed-case\">MtrE<\/jats:styled-content> and <jats:styled-content style=\"fixed-case\">MtrE<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">E<\/jats:styled-content>434K vancomycin\u2010insensitive; imply that the pump\u2013component interactions are preserved, and that the complex is stable in the absence of proton flux, thus sealing the open end of <jats:styled-content style=\"fixed-case\">MtrE<\/jats:styled-content>. Following the energy\u2010dependent dissociation of the tripartite complex, the <jats:styled-content style=\"fixed-case\">MtrE<\/jats:styled-content> channel is able to reseal, while <jats:styled-content style=\"fixed-case\">MtrE<\/jats:styled-content>\u2010<jats:styled-content style=\"fixed-case\">E<\/jats:styled-content>434K is unable to do so, resulting in the vancomycin\u2010sensitive phenotype. Thus, our findings suggest that opening of the <jats:styled-content style=\"fixed-case\">OMP<\/jats:styled-content> via interaction with the <jats:styled-content style=\"fixed-case\">MFP<\/jats:styled-content> is energy\u2010independent, while both drug export and complex dissociation require active proton flux.<\/jats:p>","DOI":"10.1111\/mmi.12211","type":"journal-article","created":{"date-parts":[[2013,4,9]],"date-time":"2013-04-09T07:38:41Z","timestamp":1365493121000},"page":"590-602","source":"Crossref","is-referenced-by-count":41,"title":["Tripartite efflux pumps: energy is required for dissociation, but not assembly or opening of the outer membrane channel of the pump"],"prefix":"10.1111","volume":"88","author":[{"given":"Thamarai K.","family":"Janganan","sequence":"first","affiliation":[{"name":"School of Biological and Biomedical Sciences Durham University  South Road Durham DH1 3LE UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vassiliy N.","family":"Bavro","sequence":"additional","affiliation":[{"name":"Institute of Microbiology and Infection, School of Immunity and Infection University of Birmingham  Edgbaston Birmingham B15 2TT UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li","family":"Zhang","sequence":"additional","affiliation":[{"name":"School of Biological and Biomedical Sciences Durham University  South Road Durham DH1 3LE UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria In\u00eas","family":"Borges\u2010Walmsley","sequence":"additional","affiliation":[{"name":"School of Biological and Biomedical Sciences Durham University  South Road Durham DH1 3LE UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adrian R.","family":"Walmsley","sequence":"additional","affiliation":[{"name":"School of Biological and Biomedical Sciences Durham University  South Road Durham DH1 3LE 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