Department of Biotechnology and Biomedicine, Technical University of Denmark, DK-2800 Kongens Lyngby, Denmark.
Department of Biotechnology and Biomedicine, Technical University of Denmark, DK-2800 Kongens Lyngby, Denmark.
Biochim Biophys Acta Proteins Proteom. 2020 Jun;1868(6):140392. doi: 10.1016/j.bbapap.2020.140392. Epub 2020 Feb 20.
The biological activity of a protein is regulated at many levels ranging from control of transcription and translation to post-translational modifications (PTM). Proteolytic processing is an irreversible PTM generating novel isoforms of a mature protein termed proteoforms. Proteoform dynamics is a major focus of current proteome research, since it has been associated with many pathological conditions. Mass-spectrometry (MS)-based proteomics and PTM-specific enrichment workflows have become the methods of choice to study proteoforms in vitro and in vivo. Here, we give an overview of currently available MS-based degradomics methods and outline how they can be optimally applied to study protease cleavage events. We discuss the advantages and disadvantages of selected approaches and describe state-of-the-art improvements in degradomics technologies. By introducing the concept of combinatorial degradomics, a combination of global discovery degradomics and highly sensitive targeted degradomics, we demonstrate how MS-based degradomics further evolves as a powerful tool in biomedical protease research.
蛋白质的生物活性在多个层面受到调控，从转录和翻译控制到翻译后修饰 (PTM)。蛋白水解加工是一种不可逆的 PTM，可产生成熟蛋白的新型同工型，称为蛋白亚型。蛋白亚型动力学是当前蛋白质组学研究的重点，因为它与许多病理状况有关。基于质谱 (MS) 的蛋白质组学和 PTM 特异性富集工作流程已成为体外和体内研究蛋白亚型的首选方法。在这里，我们概述了当前可用的基于 MS 的降解组学方法，并概述了如何将它们最佳应用于研究蛋白酶切割事件。我们讨论了选定方法的优缺点，并描述了降解组学技术的最新进展。通过引入组合降解组学的概念，即全局发现降解组学和高度敏感的靶向降解组学的组合，我们展示了基于 MS 的降解组学如何进一步发展成为生物医学蛋白酶研究的有力工具。
