Integrative analysis of the ubiquitin proteome isolated using Tandem Ubiquitin Binding Entities (TUBEs)
Integrative analysis of the ubiquitin proteome isolated using Tandem Ubiquitin Binding Entities (TUBEs)
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DOI：
10.1016/j.jprot.2011.12.001
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发表时间：
2012-06-06
影响因子：
3.3
通讯作者：
Rodriguez, Manuel S.
中科院分区：
文献类型：
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作者：
Lopitz-Otsoa, Fernando;Rodriguez-Suarez, Eva;Rodriguez, Manuel S.
蛋白酶体抑制剂在临床试验中的成功应用揭示了泛素蛋白酶体系统在药物开发中的潜力。已知通过泛素化的蛋白质重塑通过很大程度上未表征的机制来调节必需细胞因子的稳定性和活性。在这里，我们在非变性条件下使用串联重复泛素结合实体（TUBE），然后进行质谱分析，以研究可能导致识别潜在药物靶点的全局泛素化事件。使用这种方法，我们确定了643蛋白，包括已知和未知的泛素靶点，从人乳腺癌MCF 7细胞阿霉素处理。一致的全球细胞反应，这种遗传毒性的侮辱，细胞因子参与蛋白质合成，细胞运输，RNA转录后修饰和信号通路调节早期应激反应。这包括大分子复合物的组分，如蛋白酶体的亚基和调节剂，支持使用这种方法来表征由泛素化协调的分子相互作用网络。进一步的体外和计算机分析证实，这里鉴定的总蛋白质中有84%是泛素化的。更重要的是，已知生物标志物和药物开发靶点的丰富，强调了这种方法用于鉴定这种临床相关信息的潜力。本文是特刊的一部分，题为：蛋白质组学：临床联系。(C)2011 Elsevier BM. All rights reserved.
The successful use of proteasome inhibitors in clinical trials revealed the potential of the Ubiquitin Proteasome System for drug development. Protein remodeling through ubiquitylation is known to regulate the stability and activity of essential cellular factors through largely uncharacterized mechanisms. Here, we used Tandem repeated Ubiquitin Binding Entities (TUBEs) under non-denaturing conditions followed by mass spectrometry analysis to study global ubiquitylation events that may lead to the identification of potential drug targets. Using this approach we identified 643 proteins including known and unknown ubiquitin targets from human breast adenocarcinoma MCF7 cells treated with Adriamycin. Coherent with a global cellular response to this genotoxic insult, cellular factors identified are involved in protein synthesis, cellular transport, RNA post-transcriptional modification and signaling pathways regulating early stress responses. This includes components of large macromolecular complexes such as subunits and regulators of the proteasome, supporting the use of this method to characterize networks of molecular interactions coordinated by ubiquitylation. Further in vitro and in silico analysis confirmed that 84% of the total proteins identified here, are ubiquitylated. More importantly the enrichment of known biomarkers and targets for drug development, underlined the potential of this approach for the identification of this clinically relevant information. This article is part of a Special Issue entitled: Proteomics: The clinical link. (C) 2011 Elsevier BM. All rights reserved.