Role of the Na+-translocating NADH:quinone oxidoreductase in voltage generation and Na+ extrusion in Vibrio cholerae

被引:19
|
作者
Vorburger, Thomas [1 ]
Nedielkov, Ruslan [2 ,5 ]
Brosig, Alexander [3 ]
Bok, Eva [1 ]
Schunke, Emina [1 ]
Steffen, Wojtek [1 ,6 ]
Mayer, Sonja [4 ]
Goetz, Friedrich [4 ]
Moeller, Heiko M. [2 ,5 ]
Steuber, Julia [1 ]
机构
[1] Univ Stuttgart Hohenheim, Inst Microbiol, Garbenstr 30, D-70599 Stuttgart, Germany
[2] Univ Konstanz, Dept Chem, Univ Str 10, D-78457 Constance, Germany
[3] Univ Konstanz, Dept Biol, Univ Str 10, D-78457 Constance, Germany
[4] Univ Tubingen, Dept Microbial Genet, Interfac Inst Microbiol & Infect Med, Morgenstelle 28, D-72076 Tubingen, Germany
[5] Univ Potsdam, Inst Chem, Karl Liebknecht Str 24-25, D-14476 Potsdam, Germany
[6] Roche Diagnost GmbH, Nonnenwald 2, D-82377 Penzberg, Germany
来源
关键词
Nuclear magnetic resonance (NMR); Sodium transport; Vibrio cholerae; Respiration; Na+ homeostasis; Hypoosmotic stress; NADH-QUINONE OXIDOREDUCTASE; TRANSCRIPTIONAL REGULATOR; ESCHERICHIA-COLI; BINDING-SITE; PURIFICATION; TRANSPORT; NQR; IDENTIFICATION; ALGINOLYTICUS; MECHANISM;
D O I
10.1016/j.bbabio.2015.12.010
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
For Vibrio cholerae, the coordinated import and export of Na+ is crucial for adaptation to habitats with different osmolarities. We investigated the Na+-extruding branch of the sodium cycle in this human pathogen by in vivo Na-23-NMR spectroscopy. The Na+ extrusion activity of cells was monitored after adding glucose which stimulated respiration via the Na+-translocating NADH:quinone oxidoreductase (Na+-NQR). In a V. cholerae deletion mutant devoid of the Na+-NQR encoding genes (nqrA-F), rates of respiratory Na+ extrusion were decreased by a factor of four, but the cytoplasmic Na+ concentration was essentially unchanged. Furthermore, the mutant was impaired in formation of transmembrane voltage (Delta psi, inside negative) and did not grow under hypoosmotic conditions at pH 8.2 or above. This growth defect could be complemented by transformation with the plasmid encoded nqr operon. In an alkaline environment, Na+/H+ antiporters acidify the cytoplasm at the expense of the transmembrane voltage. It is proposed that, at alkaline pH and limiting Na+ concentrations, the Na+-NQR is crucial for generation of a transmembrane voltage to drive the import of H+ by electrogenic Na+/H+ antiporters. Our study provides the basis to understand the role of the Na+-NQR in pathogenicity of V. cholerae and other pathogens relying on this primary Na+ pump for respiration. (C) 2015 Elsevier B.V. All rights reserved.
引用
收藏
页码:473 / 482
页数:10
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