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

Role of the Na(+)-translocating NADH:quinone oxidoreductase in voltage generation and Na(+) extrusion in Vibrio cholerae.
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Na( )-转位 NADH:醌氧化还原酶在霍乱弧菌电压产生和 Na( ) 挤出中的作用

DOI:
10.1016/j.bbabio.2015.12.010
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发表时间:
2016
期刊:
Biochimica et biophysica acta
影响因子:
--
通讯作者:
Steuber
Steuber
中科院分区:
--
文献类型:
--
作者:
Vorburger;Nedielkov;Brosig;Schunke;Steffen;Möller;Steuber

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对于霍乱弧菌,Na+的协调输入和输出对于适应不同渗透压的栖息地至关重要。我们用活体~(23)Na-核磁共振波谱研究了这种人类病原体钠循环的Na~+-挤出分支。加入葡萄糖后,通过Na+转位NADH:Q氧化还原酶(Na+-NQR)刺激呼吸,监测细胞的Na+排出活性。在影音中。缺失Na+-NQR编码基因(nqrA-F)的霍乱突变株,呼吸Na+排出速率降低4倍,但胞浆Na+浓度基本不变。此外,突变体的跨膜电压(ΔΨ,内负值)的形成受到损害,在pH=8.2或以上的低渗透条件下不能生长。这一生长缺陷可以通过转化编码的nqroperon来补充。在碱性环境中,Na+/H+逆向转运蛋白以跨膜电压为代价酸化细胞质。我们认为,在碱性pH和限制Na+浓度的条件下,Na+-NQR对于产生跨膜电压以驱动电生Na+/H+逆向转运体的H+输入至关重要。我们的研究为了解Na+-NQR在V致病中的作用提供了基础。霍乱和其他病原体依靠这个主要的Na+泵进行呼吸。
ForVibrio 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 vivo23Na-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 aV. choleraedeletion 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 (ΔΨ, 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 encodednqroperon. 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 ofV. choleraeand other pathogens relying on this primary Na+pump for respiration.
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