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Combined top-down/bottom-up proteome analysis using electrokinetic-based multidimensional separations

机译:使用基于电动电动的多维分离的自上而下/自下而上的蛋白质组分析

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While direct analysis of tryptic peptides obtained from cell lysates offers great potential for the analysis of low abundance proteins, bottom-up approaches provide very limited molecular information about intact proteins, particularly towards the detection of post-translational modifications (PTMs). Top-down approaches, which target the analysis of intact proteins rather than protein digests, may be advantageous for the detection of PTMs. Furthermore, the top-down approach to protein sequence analysis using tandem MS may allow protein characterization far more efficiently than shotgun technologies. Realizing the full potential of a top-down approach requires that the processing and separation of intact proteins be brought to a similar level as that routinely achieved in shotgun proteomics.
机译:虽然从细胞裂解物中获得的胰蛋白酶肽的直接分析提供了对低丰度蛋白的分析的潜力,但自下而上的方法提供了关于完整蛋白质的非常有限的分子信息,特别是在检测翻译后修饰(PTMS)。自上而下的方法,其靶向完整蛋白质而不是蛋白质消化,可能是有利的,对PTMS检测有利。此外,使用串联MS的蛋白质序列分析的自上而下方法可以允许蛋白质表征远远超过霰弹枪技术。实现自上而下方法的全部潜力要求完整蛋白质的加工和分离被带到类似水平,因为在霰弹枪蛋白质组学中常规实现。

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