Characterization of MATE-type multidrug efflux pumps from Klebsiella pneumoniae MGH78578

Wakano Ogawa, Yusuke Minato, Hayata Dodan, Motoyasu Onishi, Tomofusa Tsuchiya, Teruo Kuroda

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

We previously described the cloning of genes related to drug resistance from Klebsiella pneumoniae MGH78578. Of these, we identified a putative gene encoding a MATE-type multidrug efflux pump, and named it ketM. Escherichia coli KAM32 possessing ketM on a plasmid showed increased minimum inhibitory concentrations for norfloxacin, ciprofloxacin, cefotaxime, acriflavine, Hoechst 33342, and 4′,6-diamidino-2-phenyl indole (DAPI). The active efflux of DAPI was observed in E. coli KAM32 possessing ketM on a plasmid. The expression of mRNA for ketM was observed in K. pneumoniae cells, and we subsequently disrupted ketM in K. pneumoniae ATCC10031. However, no significant changes were observed in drug resistance levels between the parental strain ATCC10031 and ketM disruptant, SKYM. Therefore, we concluded that KetM was a multidrug efflux pump, that did not significantly contribute to intrinsic resistance to antimicrobial chemicals in K. pneumoniae. MATE-type transporters are considered to be secondary transporters; therefore, we investigated the coupling cations of KetM. DAPI efflux by KetM was observed when lactate was added to produce a proton motive force, indicating that KetM effluxed substrates using a proton motive force. However, the weak efflux of DAPI by KetM was also noted when NaCl was added to the assay mixture without lactate. This result suggests that KetM may utilize proton and sodium motive forces.

Original languageEnglish
Article numbere0121619
JournalPLoS One
Volume10
Issue number3
DOIs
Publication statusPublished - Mar 25 2015

Fingerprint

Klebsiella pneumoniae
indoles
Proton-Motive Force
transporters
Pumps
Protons
drug resistance
Drug Resistance
Escherichia coli
lactates
Lactic Acid
plasmids
Plasmids
Acriflavine
acriflavine
norfloxacin
cefotaxime
Norfloxacin
Gene encoding
Cefotaxime

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Ogawa, W., Minato, Y., Dodan, H., Onishi, M., Tsuchiya, T., & Kuroda, T. (2015). Characterization of MATE-type multidrug efflux pumps from Klebsiella pneumoniae MGH78578. PLoS One, 10(3), [e0121619]. https://doi.org/10.1371/journal.pone.0121619

Characterization of MATE-type multidrug efflux pumps from Klebsiella pneumoniae MGH78578. / Ogawa, Wakano; Minato, Yusuke; Dodan, Hayata; Onishi, Motoyasu; Tsuchiya, Tomofusa; Kuroda, Teruo.

In: PLoS One, Vol. 10, No. 3, e0121619, 25.03.2015.

Research output: Contribution to journalArticle

Ogawa, W, Minato, Y, Dodan, H, Onishi, M, Tsuchiya, T & Kuroda, T 2015, 'Characterization of MATE-type multidrug efflux pumps from Klebsiella pneumoniae MGH78578', PLoS One, vol. 10, no. 3, e0121619. https://doi.org/10.1371/journal.pone.0121619
Ogawa, Wakano ; Minato, Yusuke ; Dodan, Hayata ; Onishi, Motoyasu ; Tsuchiya, Tomofusa ; Kuroda, Teruo. / Characterization of MATE-type multidrug efflux pumps from Klebsiella pneumoniae MGH78578. In: PLoS One. 2015 ; Vol. 10, No. 3.
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