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首页> 外文期刊>Biochemistry >ExbB Protein in the Cytoplasmic Membrane of Escherichia coli Forms a Stable Oligomer
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ExbB Protein in the Cytoplasmic Membrane of Escherichia coli Forms a Stable Oligomer

机译：大肠杆菌的细胞质膜中的ExbB蛋白形成稳定的寡聚物

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摘要

In Gram-negative bacteria like Escherichia coli the ExbB-ExbD-TonB protein complex isnanchored to the cytoplasmic membrane and is involved in energization of outer membrane transport. Outernmembrane proteins catalyze energy-coupled transport of scarce nutrients. Energy is derived from thenprotonmotive force of the cytoplasmic membrane which is transferred through ExbB-ExbD-TonB tonthe outer membrane transporters. Earlier studies showed that ExbB is the most abundant protein of thenExbB-ExbD-TonB complex and stabilizes TonB and ExbD. To advance understanding of the role of ExbBnin the membrane organization of the ExbB-ExbD-TonB complex, His-tagged ExbB was solubilized withndecyl maltoside and purified to electrophoretic homogeneity. Its size and shape were determined by bluennative gel electrophoresis, size exclusion chromatography, transmission electronmicroscopy, and small-anglenX-ray scattering. Decyl maltoside bound to ExbB was quantified by the anthrone method that determinesnthe sugar moiety of decyl maltoside. The results obtained with the four methods consistently indicated thatnisolated ExbB adopts a stable homooligomer with four to six monomers. We propose that the ExbBnhomooligomer forms a platform on which ExbD and TonB are assembled to form the energy-transducingncomplex in the cytoplasmic membrane.
机译：在诸如大肠杆菌的革兰氏阴性细菌中，ExbB-ExbD-TonB蛋白复合物被锚定在细胞质膜上，并参与外膜转运的能量。外膜蛋白催化稀有养分的能量耦合运输。能量来自于质膜的质子动力，质子动力通过ExbB-ExbD-TonB传递至外膜转运蛋白。早期的研究表明，ExbB是当时的ExbB-ExbD-TonB复合体中最丰富的蛋白质，可稳定TonB和ExbD。为了进一步了解ExbBn在ExbB-ExbD-TonB复合物的膜组织中的作用，将带有His标签的ExbB用十二烷基麦芽糖苷溶解并纯化至电泳均质。它的大小和形状通过增色凝胶电泳，尺寸排阻色谱，透射电镜和小角度X射线散射测定。通过确定蒽基麦芽糖苷糖部分的蒽酮方法对结合到ExbB上的癸基麦芽糖苷进行定量。四种方法获得的结果一致表明，分离出的ExbB采用具有四到六个单体的稳定的均聚物。我们提出ExbBnhomooligomer形成一个平台，在其上组装ExbD和TonB以在细胞质膜中形成能量传导复合物。

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  • 来源
    《Biochemistry》 |2010年第40期|p.8721-8728|共8页
  • 作者

    Avijit Pramanik Fajun Zhang Heinz Schwarz Frank Schreiber and Volkmar Braun;

  • 作者单位

    ‡Max Planck Institute for Developmental Biology, Spemannstrasse 35, 72076 T€ ubingen, Germany, and §Institut f€ ur AngewandtePhysik, Universit€ at T€ ubingen, Auf der Morganstelle 10, 72076 T€ ubingen, Germany;

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