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首页> 外文期刊>Journal of Molecular Biology >M13 procoat protein insertion into YidC and SecYEG proteoliposomes and liposomes.
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M13 procoat protein insertion into YidC and SecYEG proteoliposomes and liposomes.

机译：M13 procoat蛋白插入YidC和SecYEG蛋白脂质体和脂质体中。

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摘要

M13 procoat protein was one of the first model proteins used to study bacterial membrane protein insertion. It contains a signal peptide of 23 amino acid residues and is not membrane targeted by the signal recognition particle. The translocation of its periplasmic domain is independent of the preprotein translocase (SecAYEG) but requires electrochemical membrane potential and the membrane insertase YidC of Escherichia coli. We show here that YidC is sufficient for efficient membrane insertion of the purified M13 procoat protein into energized YidC proteoliposomes. When no membrane potential is applied, the insertion is substantially reduced. Only in the presence of YidC, membrane insertion occurs if bilayer integrity is preserved and membrane potential is stable for more than 20 min. A mutant of the M13 procoat protein, H5EE, with two additional negatively charged residues in the periplasmic domain inserted into YidC proteoliposomes and SecYEG proteoliposomes with equal efficiencies. We conclude that the protein can use both the YidC-only pathway and the Sec pathway. This poses the questions of how procoat H5EE is inserted in vivo and how insertion pathways are selected in the cell.
机译：M13 procoat蛋白是最早用于研究细菌膜蛋白插入的模型蛋白之一。它包含一个23个氨基酸残基的信号肽，并且未被信号识别颗粒膜靶向。其周质结构域的易位不依赖于前蛋白转位酶（SecAYEG），但需要电化学膜电位和大肠杆菌的膜插入酶YidC。我们在这里显示，YidC足以将纯化的M13 procoat蛋白有效地插入激励的YidC蛋白脂质体中。当没有施加膜电势时，插入大大减少。仅在存在YidC的情况下，如果保留双层完整性并且膜电势稳定超过20分钟，就会发生膜插入。 M13外壳蛋白的突变体H5EE，在等周质域中具有两个附加的带负电荷的残基，以相同的效率插入YidC蛋白脂质体和SecYEG蛋白脂质体。我们得出的结论是，蛋白质可以同时使用仅YidC途径和Sec途径。这提出了如何在体内插入Procoat H5EE以及如何在细胞中选择插入途径的问题。

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