Global topology analysis of the Escherichia coli inner membrane proteome
Global topology analysis of the Escherichia coli inner membrane proteome
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DOI：
10.1126/science.1109730
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发表时间：
2005-05-27
期刊：
影响因子：
56.9
通讯作者：
von Heijne, G
中科院分区：
文献类型：
--
作者：
Daley, DO;Rapp, M;von Heijne, G
众所周知，细胞膜的蛋白质补体对标准蛋白质组学分析和结构研究具有抗性。因此，膜蛋白质组仍然不明确。在这里，我们报告了一个全球性的拓扑结构分析的大肠杆菌内膜蛋白质组。利用碱性磷酸酶和绿色荧光蛋白标记的C末端，我们确定了601个内膜蛋白C末端的周质或胞质位置。通过用这些数据约束拓扑预测算法，我们得到了601种蛋白质的高质量拓扑模型，为将来对这种蛋白质组和其他膜蛋白质组的功能研究提供了坚实的基础。我们还估计了397个绿色荧光蛋白融合体的过表达潜力;结果表明，所有内膜蛋白中的很大一部分可以产生足够数量的生化和结构工作。
The protein complement of cellular membranes is notoriously resistant to standard proteomic analysis and structural studies. As a result, membrane proteomes remain ill-defined. Here, we report a global topology analysis of the Escherichia coli inner membrane proteome. Using C-terminal tagging with the alkaline phosphatase and green fluorescent protein, we established the periplasmic or cytoplasmic locations of the C termini for 601 inner membrane proteins. By constraining a topology prediction algorithm with this data, we derived high-quality topology models for the 601 proteins, providing a firm foundation for future functional studies of this and other membrane proteomes. We also estimated the overexpression potential for 397 green fluorescent protein fusions; the results suggest that a large fraction of all inner membrane proteins can be produced in sufficient quantities for biochemical and structural work.