Department of Biochemistry , University of Alberta , Edmonton , Alberta Canada.
ACS Chem Biol. 2019 Nov 15;14(11):2361-2371. doi: 10.1021/acschembio.9b00398. Epub 2019 Aug 15.
Proteases are key regulators of vital biological processes, such as apoptosis, cell differentiation, viral infection and neurodegeneration. Proteases are tightly regulated, largely because proteolysis is a unique post-translational modification (PTM) that is essentially irreversible. In order to understand the role of proteases in health and disease, the identification of protease substrates is an important step toward our understanding of their biological functions. Classic approaches for the study of proteolysis in complex mixtures employ gel electrophoresis and mass spectrometry. Such approaches typically identify a few protein substrates at a time but often fail to identify specific cleavage site locations. In contrast, modern proteomic methods using enrichment of proteolytic protein fragments can lead to the identification of hundreds of modified peptides with precise cleavage site determination in a single experiment. In this manuscript, we will review recent advances in N-terminomics methods and highlight key studies that have taken advantage of these technologies to advance our understanding of the role of proteases in cellular physiology.
蛋白酶是许多重要生命过程的关键调控因子，如细胞凋亡、细胞分化、病毒感染和神经退行性疾病等。蛋白酶的调控非常严格，这主要是因为蛋白水解是一种独特的翻译后修饰（PTM），基本上是不可逆的。为了了解蛋白酶在健康和疾病中的作用，鉴定蛋白酶的底物是理解其生物学功能的重要步骤。在复杂混合物中研究蛋白水解的经典方法采用凝胶电泳和质谱法。此类方法通常一次只能鉴定少数几种蛋白质底物，但往往无法确定特定的切割位点位置。相比之下，使用蛋白酶切蛋白片段富集的现代蛋白质组学方法可在单个实验中鉴定数百种具有精确切割位点的修饰肽。在本文中，我们将回顾 N-端组学方法的最新进展，并重点介绍利用这些技术来深入了解蛋白酶在细胞生理学中的作用的关键研究。
