Proteomic analyses reveal divergent ubiquitylation site patterns in murine tissues.
Proteomic analyses reveal divergent ubiquitylation site patterns in murine tissues.
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DOI：
10.1074/mcp.m112.017905
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发表时间：
2012-12
期刊：
影响因子：
--
通讯作者：
Choudhary C
中科院分区：
文献类型：
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作者：
Wagner SA;Beli P;Weinert BT;Schölz C;Kelstrup CD;Young C;Nielsen ML;Olsen JV;Brakebusch C;Choudhary C
蛋白质的翻译后修饰增加了细胞蛋白质组的复杂性，并使蛋白质功能能够响应环境变化而快速调节。蛋白质泛素化是一种重要的翻译后修饰，控制着蛋白质的降解、DNA损伤修复和先天免疫反应等多种生物学过程。在这里，我们结合联合收割机高分辨率质谱与单步免疫富集的双甘氨酸修饰肽映射内源性推定的泛素化位点在小鼠组织。我们在参与不同生物过程的蛋白质上鉴定了20，000多个独特的泛素化位点。我们的数据表明，泛素化调节核心信号通路共同为每个研究组织。此外，我们发现泛素化调节组织特异性信号网络。许多组织特异性的泛素化位点从大脑中获得，突出了这个器官的复杂性和独特的生理学。我们进一步证明，不同的双甘氨酸-赖氨酸特异性单克隆抗体表现出序列偏好，并且它们的互补使用增加了泛素化位点分析的深度，从而提供了一个更公正的蛋白质泛素化的观点。
Posttranslational modifications of proteins increase the complexity of the cellular proteome and enable rapid regulation of protein functions in response to environmental changes. Protein ubiquitylation is a central regulatory posttranslational modification that controls numerous biological processes including proteasomal degradation of proteins, DNA damage repair and innate immune responses. Here we combine high-resolution mass spectrometry with single-step immunoenrichment of di-glycine modified peptides for mapping of endogenous putative ubiquitylation sites in murine tissues. We identify more than 20,000 unique ubiquitylation sites on proteins involved in diverse biological processes. Our data reveals that ubiquitylation regulates core signaling pathways common for each of the studied tissues. In addition, we discover that ubiquitylation regulates tissue-specific signaling networks. Many tissue-specific ubiquitylation sites were obtained from brain highlighting the complexity and unique physiology of this organ. We further demonstrate that different di-glycine-lysine-specific monoclonal antibodies exhibit sequence preferences, and that their complementary use increases the depth of ubiquitylation site analysis, thereby providing a more unbiased view of protein ubiquitylation.