Multiple polypeptide forms observed in two-dimensional gels of Methylococcus capsulatus (Bath) polypeptides are generated during the separation procedure
Multiple polypeptide forms observed in two-dimensional gels of Methylococcus capsulatus (Bath) polypeptides are generated during the separation procedure
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DOI：
10.1002/elps.200390091
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发表时间：
2003-02-01
期刊：
影响因子：
2.9
通讯作者：
Jensen, HB
中科院分区：
文献类型：
--
作者：
Berven, FS;Karlsen, OA;Jensen, HB
我们检查了革兰氏阴性菌胶囊甲氧西林球菌(Bath)多肽的双向电泳(2-DE)凝胶图谱，发现与其他革兰氏阴性细菌多肽的2-DE凝胶中经常报道的一样，分布广泛的斑点序列。其中一些。在2-DE分离过程中，从外膜和可溶性蛋白质组分都产生了一系列多肽，而不是通过共价翻译后修饰产生的。当重新运行单个多肽点时，发现这些序列重新生成。根据Guruprasad等人的不稳定指数，分析给出这类列车的多肽都被归类为稳定多肽。(蛋白质工程)1990、4、155-161)。这种现象很可能反映了所用实验条件下多肽的构象平衡，这是标准2-DE程序的一个明显缺陷。使得凝胶图像分析变得不必要地复杂。
We have examined two-dimensional electrophoresis (2-DE) gel maps of polypeptides from the Gram-negative bacterium Methylococcus capsulatus (Bath) and found the same widespread trains of spots as often reported in 2-DE gels of polypeptides of other Gram-negative bacteria. Some of the. trains of polypeptides, both from the outer membrane and soluble protein fraction, were shown to be generated during the separation procedure of 2-DE, and not by covalent post-translational modifications. The trains were found to be regenerated when rerunning individual polypeptide spots. The polypeptides analysed giving this type of trains were all found to be classified as stable polypeptides according to the instability index of Guruprasad et al. (Protein Eng. 1990, 4, 155-161). The phenomenon most likely reflects conformational equilibria of polypeptides arising from the experimental conditions used, and is a clear drawback of the standard 2-DE procedure,. making the gel picture unnecessarily complex to analyse.