Proteome analysis of Escherichia coli K-12 by two-dimensional native-state chromatography and MALDI-MS
Proteome analysis of Escherichia coli K-12 by two-dimensional native-state chromatography and MALDI-MS
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DOI：
10.1046/j.1365-2958.2003.03294.x
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发表时间：
2003-01-01
影响因子：
3.6
通讯作者：
Hu, JC
中科院分区：
文献类型：
--
作者：
Champion, MM;Campbell, CS;Hu, JC
为了鉴定大肠杆菌K-12 MG 1655在限定培养基中指数生长期间表达的蛋白质，我们分离了大肠杆菌的可溶性蛋白质。大肠杆菌在二维的天然状态的高效液相色谱，并检查了蛋白质混合物的组成部分，在每一个380肽质量指纹图谱。到目前为止，我们已经确定了310个基因的产品，涵盖了广泛的细胞功能。通过将蛋白质分配至特定的第一维级分与通过二维凝胶电泳可视化的蛋白质进行比较来进行蛋白质分配的验证。蛋白质的共分馏表明多蛋白复合物的组分可能是同一的。这种方法产生了高通量的蛋白质凝胶独立鉴定。它也可以用来分配身份的斑点可视化的二维凝胶，并应是有用的，以评估菌株之间或不同的生理状态之间的差异表达的蛋白质组含量和蛋白质复合物。
To identify proteins expressed in Escherichia coli K-12 MG1655 during exponential growth in defined medium, we separated soluble proteins of E. coli over two dimensions of native-state high-performance liquid chromatography, and examined the components of the protein mixtures in each of 380 fractions by peptide mass fingerprinting. To date, we have identified the products of 310 genes covering a wide range of cellular functions. Validation of protein assignments was made by comparing the assignments of proteins to specific first-dimension fractions to proteins visualized by two-dimensional gel electrophoresis. Co-fractionation of proteins suggests the possible identities of components of multiprotein complexes. This approach yields high-throughput gel-independent identification of proteins. It can also be used to assign identities to spots visualized by two-dimensional gels, and should be useful to evaluate differences in expressed proteome content and protein complexes among strains or between different physiological states.