The Na+- translocating NADH: Quinone oxidoreductase enhances oxidative stress in the cytoplasm of Vibrio cholerae

Valentin Muras, Paul Dogaru-Kinn, Yusuke Minato, Claudia C. Häse, Julia Steuber

研究成果: Article査読

13 被引用数 (Scopus)

抄録

We searched for a source of reactive oxygen species (ROS) in the cytoplasm of the human pathogen Vibrio cholerae and addressed the mechanism of ROS formation using the dye 2',7'-dichlorofluorescein diacetate (DCFH-DA) in respiring cells. By comparing V. cholerae strains with or without active Na+-translocating NADH:quinone oxidoreductase (Na+-NQR), this respiratory sodium ion redox pump was identified as a producer of ROS in vivo. The amount of cytoplasmic ROS detected in V. cholerae cells producing variants of Na+-NQR correlated well with rates of superoxide formation by the corresponding membrane fractions. Membranes from wild-type V. cholerae showed increased superoxide production activity (9.8 ± 0.6 μmol superoxide min-1 mg-1 membrane protein) compared to membranes from the mutant lacking Na+-NQR (0.18±0.01 μmol min-1 mg-1). Overexpression of plasmid-encoded Na+-NQR in the nqr deletion strain resulted in a drastic increase in the formation of superoxide (42.6±2.8 μmol min-1 mg-1). By analyzing a variant of Na+-NQR devoid of quinone reduction activity, we identified the reduced flavin adenine dinucleotide (FAD) cofactor of cytoplasmic NqrF subunit as the site for intracellular superoxide formation in V. cholerae. The impact of superoxide formation by the Na+-NQR on the virulence of V. cholerae is discussed.

本文言語English
ページ(範囲)2307-2317
ページ数11
ジャーナルJournal of Bacteriology
198
17
DOI
出版ステータスPublished - 2016
外部発表はい

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology

フィンガープリント 「The Na<sup>+</sup>- translocating NADH: Quinone oxidoreductase enhances oxidative stress in the cytoplasm of Vibrio cholerae」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。

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