{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T23:57:39Z","timestamp":1791417459883,"version":"4.3.3"},"reference-count":33,"publisher":"American Society for Microbiology","issue":"9","license":[{"start":{"date-parts":[[2010,9,1]],"date-time":"2010-09-01T00:00:00Z","timestamp":1283299200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2010,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            AcrAB-TolC is the major constitutively expressed efflux pump system that provides resistance to a variety of antimicrobial agents and dyes in\n            <jats:italic>Escherichia coli<\/jats:italic>\n            . However, no systematically optimized real-time dye efflux assay has been published for the measurement of its activity and for detection of possible competition between substrates. Here, we report on the development of such an assay using a lipophilic dye, Nile Red. Energy-depleted cells were loaded with the dye in the presence of low (10 \u03bcM or less) concentrations of the proton conductor carbonyl cyanide\n            <jats:italic>m<\/jats:italic>\n            -chlorophenylhydrazone (CCCP). The CCCP was then removed, and efflux was triggered by energization with glucose. Various known efflux pump inhibitors and antimicrobials were checked for the ability to slow down Nile Red efflux, presumably through competition. Besides the known inhibitors Phe-Arg-\u03b2-naphthylamide and 1-naphthyl-methylpiperazine, several tetracyclic compounds (doxorubicin, minocycline, chlortetracycline, doxycycline, and tetracycline) and tetraphenylphosphonium chloride were found to interfere with dye efflux. This inhibition could not be explained by the depletion of proton motive force. None of the other tested antimicrobials, including macrolides, fluoroquinolones, and \u03b2-lactams, had any impact on Nile Red efflux, even at concentrations of up to 1 mM.\n          <\/jats:p>","DOI":"10.1128\/aac.00620-10","type":"journal-article","created":{"date-parts":[[2010,7,7]],"date-time":"2010-07-07T02:37:20Z","timestamp":1278470240000},"page":"3770-3775","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":147,"title":["Optimized Nile Red Efflux Assay of AcrAB-TolC Multidrug Efflux System Shows Competition between Substrates"],"prefix":"10.1128","volume":"54","author":[{"given":"Ju\u0308rgen A.","family":"Bohnert","sequence":"first","affiliation":[{"name":"Department of Molecular and Cell Biology, University of California, Berkeley, California 94720-3202"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brian","family":"Karamian","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cell Biology, University of California, Berkeley, California 94720-3202"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiroshi","family":"Nikaido","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cell Biology, University of California, Berkeley, California 94720-3202"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.187.6.1923-1929.2005"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.49.2.849-852.2005"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00912-08"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.01048-06"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-2275(20)34307-8"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ab.2009.07.001"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.45.5.1467-1472.2001"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.mi.39.100185.001251"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.44.4.814-820.2000"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.179.7.2189-2193.1997"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1038\/35016007"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.01714-09"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.45.1.105-116.2001"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.str.2005.11.015"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature05076"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature01050"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M308893200"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0901695106"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1369-5274(98)80083-0"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.183.20.5803-5812.2001"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.184.8.2319-2323.2002"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.272.35.21964"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.178.1.306-308.1996"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.1128\/jb.127.3.1188-1196.1976"},{"key":"e_1_3_2_26_2","doi-asserted-by":"publisher","DOI":"10.1021\/jm9904598"},{"key":"e_1_3_2_27_2","doi-asserted-by":"publisher","DOI":"10.1002\/jss.400070104"},{"key":"e_1_3_2_28_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1131542"},{"key":"e_1_3_2_29_2","first-page":"e7","volume":"5","year":"2007","unstructured":"Sennhauser, G., P. Amstutz, C. Briand, O. Storchenegger, and M. G. Grutter. 2007. Drug export pathway of multidrug exporter AcrB revealed by DARPin inhibitors. PLoS. Biol.5:e7.","journal-title":"PLoS. 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