Secondary-site mutation restores the transport defect caused by the transmembrane domain mutation of the xenobiotic transporter MexB in Pseudomonas aeruginosa

Hiroshi Yoneyama, Hideaki Maseda, Taka aki Yamabayashi, Shotaro Izumi, Taiji Nakae

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3 Citations (Scopus)

Abstract

It has been suggested that the MexB subunit of the MexAB-OprM efflux transporter of Pseudomonas aeruginosa exports xenobiotics in an energy-dependent manner. To investigate the role of the transmembrane segments (TMS) of MexB in the transporter activity, we isolated 24 spontaneous mutants showing hypersusceptibility to antibiotics. Among them, three mutations were located at TMS-3, TMS-4, and TMS-10 having amino acid substitution Leu376-vPro, Gly397vVal, and Val928vGly, respectively. A secondary mutation, which suppressed the defect caused by the Val928vGly mutation in TMS-10, was found at the 403rd amino acid residue in TMS-4 with a change of glycine to serine, suggesting that TMS-4 and TMS-10 may be in close proximity. This result provided strong support for the recent notion that negatively charged residues in TMS-4 might form a salt-bridge with a positive charge in TMS-10 (Guan, L., and Nakae, T. (2001) J. Bacteriol. 183, 1734-1739). The transporter function impaired by the Gly397vVal mutation in TMS-4 was recovered by the secondary mutation, Gln998vHis, in the loop between TMS-11 and TMS-12, thereby suggesting that TMS-4 and TMS-11 or TMS-12 might also be in close proximity. Thus, it is most likely that TMS-4, TMS-10, and TMS-11 or TMS-12 are packed close three dimensionally.

Original languageEnglish
Pages (from-to)513-518
Number of pages6
JournalBiochemical and biophysical research communications
Volume292
Issue number2
DOIs
Publication statusPublished - 2002
Externally publishedYes

Keywords

  • Antibiotic
  • Drug resistance
  • Efflux transporter
  • MDR
  • Membrane protein
  • Transmembrane segment
  • Xenobiotics

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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