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首页> 外文期刊>Journal of bacteriology >Functional Expression in Escherichia coli and Membrane Topology of Porin HopE, a Member of a Large Family of Conserved Proteins in Helicobacter pylori
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Functional Expression in Escherichia coli and Membrane Topology of Porin HopE, a Member of a Large Family of Conserved Proteins in Helicobacter pylori

机译：大肠杆菌中的功能表达和Porin HopE（幽门螺杆菌中一个大的保守蛋白家族成员）的膜拓扑结构

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摘要

HopE is one of the smallest members of a family of 31 outer membrane proteins in Helicobacter pylori and has been shown to function as a porin. In this study it was cloned intoEscherichia coli where it was expressed in the outer membrane, as confirmed by indirect immunofluorescence using HopE-specific antibodies. HopE purified from E. colireconstituted channels in planar bilayer membranes that were the same size as those formed by HopE purified from H. pylori. A model of the membrane topology of HopE was constructed and indicated that this protein formed a β-barrel with 16 transmembrane amphipathic β-strands. The accuracy of this model was tested by linker insertion mutagenesis, assuming that, like other porins, amino acid insertions were not tolerated in the transmembrane β-strands but were tolerated in the adjoining loop regions. Generally, the results obtained with a series of 12 insertions of the sequence RSKDV and two substitutions were consistent with the topological model. The preponderance of amino acids that were conserved in the extended family of HopE paralogs were predicted to be within the membrane and comprised 45% of all residues in the membrane.
机译：HopE是幽门螺杆菌（Helicobacter pylori）中31种外膜蛋白家族中最小的成员之一，已被证明具有孔蛋白的功能。在这项研究中，将其克隆到大肠埃希菌中，并在外膜中表达，这是通过使用HopE特异性抗体进行的间接免疫荧光证实的。从 E中纯化的HopE。平面双层膜中的大肠杆菌重构通道，其大小与从 H中纯化的HopE形成的通道相同。幽门螺旋杆菌。构建了HopE膜拓扑结构模型，表明该蛋白形成了带有16个跨膜两亲性β链的β桶。该模型的准确性通过接头插入诱变测试，假设像其他孔蛋白一样，氨基酸插入在跨膜β链中不被接受，但在相邻的环区域中被接受。通常，使用序列RSKDV的12个插入序列和两次替换获得的结果与拓扑模型一致。预计在HopE旁系同源物扩展家族中保守的氨基酸主要在膜内，占膜中所有残基的45％。

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