Hirata Takahiro, Saito Asami, Nishino Kunihiko, Tamura Norihisa, Yamaguchi Akihito
Department of Cell Membrane Biology, Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki-shi, Osaka 567-0047, Japan.
Antimicrob Agents Chemother. 2004 Jun;48(6):2179-84. doi: 10.1128/AAC.48.6.2179-2184.2004.
The activity of tigecycline, 9-(t-butylglycylamido)-minocycline, against Escherichia coli KAM3 (acrB) strains harboring plasmids encoding various tetracycline-specific efflux transporter genes, tet(B), tet(C), and tet(K), and multidrug transporter genes, acrAB, acrEF, and bcr, was examined. Tigecycline showed potent activity against all three Tet-expressing, tetracycline-resistant strains, with the MICs for the strains being equal to that for the host strain. In the Tet(B)-containing vesicle study, tigecycline did not significantly inhibit tetracycline efflux-coupled proton translocation and at 10 microM did not cause proton translocation. This suggests that tigecycline is not recognized by the Tet efflux transporter at a low concentration; therefore, it exhibits significant antibacterial activity. These properties can explain its potent activity against bacteria with a Tet efflux resistance determinant. Tigecycline induced the Tet(B) protein approximately four times more efficiently than tetracycline, as determined by Western blotting, indicating that it is at least recognized by a TetR repressor. The MICs for multidrug efflux proteins AcrAB and AcrEF were increased fourfold. Tigecycline inhibited active ethidium bromide efflux from intact E. coli cells overproducing AcrAB. Therefore, tigecycline is a possible substrate of AcrAB and its close homolog, AcrEF, which are resistance-modulation-division-type multicomponent efflux transporters.
对携带编码各种四环素特异性外排转运蛋白基因(tet(B)、tet(C)和tet(K))以及多药转运蛋白基因(acrAB、acrEF和bcr)的质粒的大肠杆菌KAM3(acrB)菌株,检测了替加环素(9-(叔丁基甘氨酰胺)-米诺环素)的活性。替加环素对所有三种表达Tet的四环素耐药菌株均显示出强效活性,这些菌株的最低抑菌浓度(MIC)与宿主菌株相同。在含Tet(B)的囊泡研究中,替加环素未显著抑制四环素外排偶联的质子转运,在10 μM时也未引起质子转运。这表明替加环素在低浓度下不被Tet外排转运蛋白识别;因此,它表现出显著的抗菌活性。这些特性可以解释其对具有Tet外排耐药决定簇的细菌的强效活性。通过蛋白质免疫印迹法测定,替加环素诱导Tet(B)蛋白的效率比四环素高约四倍,表明它至少被TetR阻遏物识别。多药外排蛋白AcrAB和AcrEF的MIC增加了四倍。替加环素抑制了过量表达AcrAB的完整大肠杆菌细胞中溴化乙锭的主动外排。因此,替加环素可能是AcrAB及其紧密同源物AcrEF的底物,它们是耐药调节分裂型多组分外排转运蛋白。