He Gui-Xin, Kuroda Teruo, Mima Takehiko, Morita Yuji, Mizushima Tohru, Tsuchiya Tomofusa
Department of Microbiology, Faculty of Pharmaceutical Sciences, Okayama University, Tsushima, Okayama, 700-8530, Japan.
J Bacteriol. 2004 Jan;186(1):262-5. doi: 10.1128/JB.186.1.262-265.2004.
We cloned the gene PA1361 (we designated the gene pmpM), which seemed to encode a multidrug efflux pump belonging to the MATE family, of Pseudomonas aeruginosa by the PCR method using the drug-hypersensitive Escherichia coli KAM32 strain as a host. Cells of E. coli possessing the pmpM gene showed elevated resistance to several antimicrobial agents. We observed energy-dependent efflux of ethidium from cells possessing the pmpM gene. We found that PmpM is an H(+)-drug antiporter, and this finding is the first reported case of an H(+)-coupled efflux pump in the MATE family. Disruption and reintroduction of the pmpM gene in P. aeruginosa revealed that PmpM is functional and that benzalkonium chloride, fluoroquinolones, ethidium bromide, acriflavine, and tetraphenylphosphonium chloride are substrates for PmpM in this microorganism.
我们通过PCR方法,以药物敏感型大肠杆菌KAM32菌株为宿主,克隆了铜绿假单胞菌中似乎编码属于多药及毒性化合物外排(MATE)家族的多药外排泵的基因PA1361(我们将该基因命名为pmpM)。携带pmpM基因的大肠杆菌细胞对几种抗菌剂的耐药性增强。我们观察到携带pmpM基因的细胞中存在能量依赖的溴化乙锭外排。我们发现PmpM是一种H(+)-药物反向转运体,这一发现是MATE家族中首次报道的H(+)-偶联外排泵的案例。在铜绿假单胞菌中对pmpM基因进行破坏和重新导入,结果表明PmpM具有功能,并且在这种微生物中,苯扎氯铵、氟喹诺酮类、溴化乙锭、吖啶黄素和氯化四苯基鏻是PmpM的底物。