蛋白质组学是全景式鉴定、定量蛋白质,并研究蛋白质功能的学科。基于高分辨质谱的鸟枪法蛋白组学研究技术首先利用不同的位点特异性蛋白酶对复杂蛋白质样品进行酶解,进而利用质谱获得蛋白质相关的定性和定量信息。为了获得高质量的质谱信息,前期的样品处理和质谱数据采集同样重要。本文对蛋白组学中常用的Trypsin、Lys-C、Glu-C等位点特异性蛋白酶的酶切特点进行了总结,并综述了目前常用的几种酶切组合策略和样本预处理技术在提高蛋白质组学研究效率中的作用。
Proteomics is a powerful subject focusing on large-scale study of protein structures and functions. A complete enzymatic digestion of protein complexes is the key step in modern high-resolution and high-throughput mass spectrometry(MS)-based identification and quantification. To achieve MS analysis, both peptide sample pretreatment and data acquisition are prerequisite in proteomic studies. In this paper, we summarized both the enzymatic proprieties of three common proteolytic enzymes, Trypsin, Lys-C and Glu-C, the optimization of multi-enzyme combination and an advanced sample pretreatment in proteomics research.