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首页> 外文期刊>Journal of proteome research >Integration of multidimensional chromatographic protein separations with a combined 'top-down' and 'bottom-up' proteomic strategy.
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Integration of multidimensional chromatographic protein separations with a combined 'top-down' and 'bottom-up' proteomic strategy.

机译：多维色谱蛋白质分离与“自上而下”和“自下而上”的蛋白质组学策略相结合。

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摘要

In this paper, we present a combined top-down/bottom-up proteomic analysis workflow for the characterization of proteomic samples. This workflow combines protein fractionation (multidimensional chromatographic separation) with parallel online ESI-TOF-MS intact protein analysis, and fraction collection. Collected fractions were digested and protein identifications were produced using MALDI Q-TOF-MS analysis. These identifications were then linked with corresponding ESI-TOF-MS intact protein mass data to permit full protein characterization. This methodology was applied to an E. coli cytosolic protein fraction, and enabled the identification and characterization of proteins exhibiting co-translational processing, post-translational modification, and proteolytic processing events. The approach also provided the ability to distinguish between closely related protein isoforms. The summary of results from this study indicated that roughly one-third of all detected components generated corresponding data fromboth top-down and bottom-up analyses, and that significant and novel information can be derived from this application of the hybrid analytical methodology.
机译：在本文中，我们提出了一种自上而下/自下而上的蛋白质组学分析工作流程，用于蛋白质组样品的表征。该工作流程将蛋白质分离（多维色谱分离）与并行在线ESI-TOF-MS完整蛋白质分析和馏分收集结合在一起。消化收集的馏分，并使用MALDI Q-TOF-MS分析进行蛋白质鉴定。然后将这些鉴定与相应的ESI-TOF-MS完整蛋白质质量数据关联起来，以进行完整的蛋白质表征。此方法应用于大肠杆菌胞质蛋白部分，并能够鉴定和表征表现出共翻译过程，翻译后修饰和蛋白水解过程事件的蛋白质。该方法还提供了区分紧密相关的蛋白质同工型的能力。这项研究的结果摘要表明，从上至下和自下而上的分析中，所有检测到的成分中大约有三分之一生成了相应的数据，并且可以从此混合分析方法的应用中得出重要而新颖的信息。

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