Proteomic approaches for the profiling of ubiquitylation events and their applications in drug discovery.
Proteomic approaches for the profiling of ubiquitylation events and their applications in drug discovery.
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DOI：
10.1016/j.jprot.2020.103996
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发表时间：
2020-10
影响因子：
3.3
通讯作者：
Zhanhong Hu;Hongcheng Li;Xiaohui Wang;K. Ullah;Guoqiang Xu
中科院分区：
文献类型：
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作者：
Zhanhong Hu;Hongcheng Li;Xiaohui Wang;K. Ullah;Guoqiang Xu
在真核细胞中，蛋白质泛素化几乎调控着基因表达、DNA修复、细胞增殖和凋亡等生物学过程的各个方面。由泛素系统中酶的异常表达引起的蛋白质泛素化的失调导致许多疾病的发作，包括癌症、神经退行性疾病和代谢综合征。因此，靶向泛素系统成为药物发现的一个很有前途的研究领域。蛋白质泛素化位点的鉴定对于揭示与疾病和特定信号通路相关的关键泛素化事件以及阐明特定泛素化事件的生物学功能至关重要。许多方法，丰富的泛素化的蛋白质和泛素化的肽在蛋白质和肽的水平已被开发，以方便他们的鉴定MS。在本文中，我们将回顾蛋白质组学方法可用于鉴定的泛素化事件在蛋白质组规模，并讨论其优点和局限性。我们还将简要介绍泛素化事件谱在药物靶点发现和蛋白水解靶向嵌合体（PROTAC）靶点验证中的应用。意义泛素化在真核细胞的许多生物学过程中起着重要的调节作用。泛素化位点的鉴定可以为特定修饰底物的功能研究提供必要的信息。由于泛素化蛋白质的丰度比非泛素化蛋白质低得多，泛素化蛋白质或肽的富集对于它们的MS鉴定至关重要。本文综述了不同的富集方法，以促进它们的分离和MS鉴定，并讨论了这些方法的优点和缺点。泛素化事件谱在药物靶点发现和未来研究方向的应用将有利于研究界。
Protein ubiquitylation regulates almost all aspects of the biological processes including gene expression, DNA repair, cell proliferation and apoptosis in eukaryotic cells. Dysregulation of protein ubiquitylation caused by abnormal expression of enzymes in the ubiquitin system results in the onset of many diseases including cancer, neurodegenerative diseases, and metabolic syndromes. Therefore, targeting the ubiquitin system becomes a promising research area in drug discovery. Identification of protein ubiquitylation sites is critical for revealing the key ubiquitylation events associated with diseases and specific signaling pathways and for elucidating the biological functions of the specific ubiquitylation events. Many approaches that enrich for the ubiquitylated proteins and ubiquitylated peptides at the protein and peptide levels have been developed to facilitate their identification by MS. In this paper, we will review the proteomic approaches available for the identification of ubiquitylation events at the proteome scale and discuss their advantages and limitations. We will also brief the application of the profiling of ubiquitylation events in drug target discovery and in target validation for proteolysis-targeting chimera (PROTAC). Possible future research directions in this field will also be discussed.SignificanceUbiquitylation plays critical roles in regulating many biological processes in eukaryotic cells. Identification of ubiquitylation sites can provide the essential information for the functional study of the specific modified substrates. Since ubiquitylated proteins have much lower abundance than non-ubiquitylated proteins, enrichment of ubiquitylated proteins or peptides is critical for their identification by MS. This review focuses on different enrichment approaches that facilitate their isolation and identification by MS and discusses the advantages and drawbacks of these approaches. The application of the profiling of ubiquitylation events in drug target discovery and future research directions will be beneficial to the research community.