Turner R J, Taylor D E, Weiner J H
Department of Biochemistry, University of Alberta, Edmonton, Canada.
Antimicrob Agents Chemother. 1997 Feb;41(2):440-4. doi: 10.1128/AAC.41.2.440.
The genes tehAB located at 32.3 min on the Escherichia coli chromosome were initially identified by their ability to mediate resistance to potassium tellurite (128 micrograms of K2TeO3 per ml) when overexpressed with a high-copy-number plasmid. The genes encode an integral membrane protein (TehA) of 36 kDa with 10 putative transmembrane segments and a second protein (TehB) of 23 kDa. Overexpression of TehAB results in hypersensitivity to dequalinium CI and methyl viologen (paraquat). Expression of TehA alone gives similar hypersensitivity. Overexpression of TehA gave resistance to tetraphenylarsonium CI, ethidium bromide, crystal violet and proflavin. The efflux of ethidium, measured by fluorescence quenching, revealed that TehA transported ethidium at twice the control rate and 10% of the rate of the highly resistant efflux transporter Emr Eco. Addition of tellurite had no effect on ethidium transport. In addition to the ethidium transport assay, a proflavin fluorescence assay which was approximately 200-fold more sensitive was also used. TehA was also found to have proflavin efflux activity. The addition of TeO32- to the proflavin transport assay on TehA caused a 20% increase in transport rate. Both ethidium and proflavin transport were found to be energy dependent.
位于大肠杆菌染色体32.3分钟处的tehAB基因最初是通过其在高拷贝数质粒过表达时介导对亚碲酸钾(每毫升128微克K2TeO3)抗性的能力而被鉴定出来的。这些基因编码一个36 kDa的整合膜蛋白(TehA),具有10个推定的跨膜区段,以及一个23 kDa的第二种蛋白(TehB)。TehAB的过表达导致对氯化喹吖因和甲基紫精(百草枯)超敏。单独表达TehA也产生类似的超敏反应。TehA的过表达赋予对氯化四苯基砷、溴化乙锭、结晶紫和原黄素的抗性。通过荧光猝灭测量溴化乙锭的外排,结果显示TehA转运溴化乙锭的速率是对照速率的两倍,是高抗性外排转运蛋白Emr Eco速率的10%。添加亚碲酸盐对溴化乙锭的转运没有影响。除了溴化乙锭转运测定外,还使用了灵敏度约高200倍的原黄素荧光测定法。还发现TehA具有原黄素外排活性。在对TehA的原黄素转运测定中添加TeO32-导致转运速率增加20%。发现溴化乙锭和原黄素的转运均依赖能量。