A Major Li+ Extrusion System NhaB of Pseudomonas aeruginosa: Comparison with the Major Na+ Extrusion System NhaP

Teruo Kuroda, Naoko Fujita, Junko Utsugi, Masayuki Kuroda, Tohru Mizushima, Tomofusa Tsuchiya

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

A gene encoding a Li+ extrusion system was cloned from the chromosomal DNA of Pseudomonas aeruginosa and expressed in Escherichia coli cells. The gene enabled growth of E. coli KNabc cells, which were unable to grow in the presence of 10 mm LiCl or 0.1 M NaCl because of the lack of major Na+(Li+)/H+ antiporters. We detected Li +/H+ and Na+/H+ antiport activities in membrane vesicles prepared from E. coli KNabc cells that harbored a plasmid carrying the cloned gene. Activity of this antiporter was pH-dependent with an optimal pH activity between pH 7.5 and 8.5. These properties indicate that this antiporter is different from NhaP, an Na+/H+ antiporter from P. aeruginosa that we reported previously, and that is rather specific to Na+ but it cannot extrude Li+ effectively. The gene was sequenced and an open reading frame (ORF) was identified. The amino acid sequence deduced from the ORF showed homology (about 60% identity and 90% similarity) with that of the NhaB Na+/H+ antiporters of E. coli and Vibrio parahaemolyticus. Thus, we designated the antiporter as NhaB of P. aeruginosa. E. coli KNabc carrying the nhaB gene from P. aeruginosa was able to grow in the presence of 10 to 50 mM LiCl, although KNabc carrying nhaP was unable to grow in these conditions. The antiport activity of NhaB from P. aeruginosa was produced in E. coli and showed apparent Km values for Li + and Na+ of 2.0 mM and 1.3 mM, respectively. The antiport activity was inhibited by amiloride with a Ki value for Li+ and Na+ of 0.03 mM and 0.04 mM, respectively.

Original languageEnglish
Pages (from-to)243-250
Number of pages8
JournalMicrobiology and Immunology
Volume48
Issue number4
Publication statusPublished - 2004
Externally publishedYes

Fingerprint

Pseudomonas aeruginosa
Escherichia coli
Antiporters
Sodium-Hydrogen Antiporter
Ion Transport
Genes
Open Reading Frames
Vibrio parahaemolyticus
Amiloride
Amino Acid Sequence
Plasmids
Membranes
DNA
Growth

Keywords

  • Antiporter
  • Li extrusion
  • Li resistance
  • NhaB
  • Pseudomans aeruginosa

ASJC Scopus subject areas

  • Immunology and Microbiology(all)
  • Microbiology (medical)
  • Microbiology

Cite this

Kuroda, T., Fujita, N., Utsugi, J., Kuroda, M., Mizushima, T., & Tsuchiya, T. (2004). A Major Li+ Extrusion System NhaB of Pseudomonas aeruginosa: Comparison with the Major Na+ Extrusion System NhaP. Microbiology and Immunology, 48(4), 243-250.

A Major Li+ Extrusion System NhaB of Pseudomonas aeruginosa : Comparison with the Major Na+ Extrusion System NhaP. / Kuroda, Teruo; Fujita, Naoko; Utsugi, Junko; Kuroda, Masayuki; Mizushima, Tohru; Tsuchiya, Tomofusa.

In: Microbiology and Immunology, Vol. 48, No. 4, 2004, p. 243-250.

Research output: Contribution to journalArticle

Kuroda, T, Fujita, N, Utsugi, J, Kuroda, M, Mizushima, T & Tsuchiya, T 2004, 'A Major Li+ Extrusion System NhaB of Pseudomonas aeruginosa: Comparison with the Major Na+ Extrusion System NhaP', Microbiology and Immunology, vol. 48, no. 4, pp. 243-250.
Kuroda, Teruo ; Fujita, Naoko ; Utsugi, Junko ; Kuroda, Masayuki ; Mizushima, Tohru ; Tsuchiya, Tomofusa. / A Major Li+ Extrusion System NhaB of Pseudomonas aeruginosa : Comparison with the Major Na+ Extrusion System NhaP. In: Microbiology and Immunology. 2004 ; Vol. 48, No. 4. pp. 243-250.
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