The Respiratory Complex I (NDH I) from Klebsiella pneumoniae, a Sodium Pump*

The Respiratory Complex I (NDH I) from Klebsiella pneumoniae, a Sodium Pump*
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肺炎克雷伯菌的呼吸复合物 I (NDH I),钠泵*

DOI:
10.1074/jbc.m204860200
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发表时间:
2002
期刊:
The Journal of Biological Chemistry
影响因子:
--
通讯作者:
J. Steuber
J. Steuber
中科院分区:
--
文献类型:
--
作者:
A. Gemperli;P. Dimroth;J. Steuber

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肺炎克雷伯菌产电的NADH:Q氧化还原酶转运Na+离子。纯化后的复合体比钠离子转运活性从天然膜泡的0.2molmolmin−1 mg−1提高到重组酶样品的4.7molmolminμ1 mg−1。在纯化过程中,对复合体I的典型辅因子进行了浓缩,得到了∼17nmol mg−1铁、24nmol mg−1酸不稳定硫化物和0.79nmol mg−1FMN。该酶含有∼1.2nmomg−1Q6和1.5nmomg−1Q8。NADH对泛醌的还原是依赖于Na+的,这表明了泵的化学和矢量反应的耦合。Na+活化谱符合Hill方程,Hill系数KH(Na+)=1.96,在0.33 mM Na+处出现半最大饱和度。重组肺炎克雷伯菌复合体I催化脱氨基-NADH氧化、Q1还原和Na+转位,比活性分别为2.6U mg−1、2.4U mg−1和4.7U mg−1,表明Na+/电子计量比为1。
The electrogenic NADH:Q oxidoreductase from the enterobacterium Klebsiella pneumoniae transports Na+ ions. The complex was purified with an increase of the specific Na+ transport activity from 0.2 μmol min−1 mg−1 in native membrane vesicles to 4.7 μmol min−1 mg−1 in reconstituted enzyme specimens. The subunit pattern resembled that of complex I fromEscherichia coli, and two prominent polypeptides were identified as the NuoF and NuoG subunits of complex I. During purification the typical cofactors of complex I were enriched to yield ∼17 nmol mg−1 iron, 24 nmol mg−1acid-labile sulfide, and 0.79 nmol mg−1 FMN in the purified sample. The enzyme contained ∼1.2 nmol mg−1 Q6 and 1.5 nmol mg−1 Q8. The reduction of ubiquinone by NADH was Na+-dependent, which indicates coupling of the chemical and the vectorial reaction of the pump. The Na+ activation profile corresponded to the Hill equation with a Hill coefficient KH(Na+) = 1.96 and with a half-maximal saturation at 0.33 mm Na+. The reconstituted complex I from Klebsiella pneumoniae catalyzed deamino-NADH oxidation, Q1 reduction, and Na+ translocation with specific activities of 2.6 units mg−1, 2.4 units mg−1, and 4.7 units mg−1, respectively, which indicate a Na+/electron stoichiometry of one.
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影响因子: --
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