Native Proteomics in Discovery Mode Using Size-Exclusion Chromatography–Capillary Zone Electrophoresis–Tandem Mass Spectrometry,Analytical Chemistry

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Native Proteomics in Discovery Mode Using Size-Exclusion Chromatography–Capillary Zone Electrophoresis–Tandem Mass Spectrometry
Analytical Chemistry ( IF 6.7 ) Pub Date : 2018-08-07 00:00:00 , DOI: 10.1021/acs.analchem.8b02725
Xiaojing Shen ₁ , Qiang Kou ₂ , Ruiqiong Guo ₁ , Zhichang Yang ₁ , Daoyang Chen ₁ , Xiaowen Liu _{2,

3} , Heedeok Hong ₁ , Liangliang Sun ₁

Affiliation

Native proteomics aims to characterize complex proteomes under native conditions and ultimately produces a full picture of endogenous protein complexes in cells. It requires novel analytical platforms for high-resolution and liquid-phase separation of protein complexes prior to native mass spectrometry (MS) and MS/MS. In this work, size-exclusion chromatography (SEC)–capillary zone electrophoresis (CZE)–MS/MS was developed for native proteomics in discovery mode, resulting in the identification of 144 proteins, 672 proteoforms, and 23 protein complexes from the Escherichia coli proteome. The protein complexes include four protein homodimers, 16 protein–metal complexes, two protein–[2Fe-2S] complexes, and one protein–glutamine complex. Half of them have not been reported in the literature. This work represents the first example of online liquid-phase separation–MS/MS for the characterization of a complex proteome under the native condition, offering the proteomics community an efficient and simple platform for native proteomics.

中文翻译：

使用尺寸排阻色谱-毛细管区带电泳-串联质谱法在发现模式下进行天然蛋白质组学

天然蛋白质组学旨在表征天然条件下的复杂蛋白质组，并最终产生细胞内源蛋白质复合物的全貌。它需要新颖的分析平台，在天然质谱 (MS) 和 MS/MS 之前对蛋白质复合物进行高分辨率和液相分离。在这项工作中，尺寸排阻色谱 (SEC)-毛细管区带电泳 (CZE)-MS/MS 被开发用于发现模式中的天然蛋白质组学，从而鉴定了来自大肠杆菌的 144 种蛋白质、672 种蛋白质形式和 23 种蛋白质复合物蛋白质组。蛋白质复合物包括四种蛋白质同二聚体、16种蛋白质-金属复合物、两种蛋白质-[2Fe-2S]复合物和一种蛋白质-谷氨酰胺复合物。其中一半未见文献报道。这项工作代表了在线液相分离 MS/MS 在天然条件下表征复杂蛋白质组的第一个例子，为蛋白质组学界提供了一个高效、简单的天然蛋白质组学平台。

更新日期：2018-08-07

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