翻译后修饰蛋白质组学研究的技术策略

 蛋白质组学早期研究的绝大部分工作是在关注细胞不同生长时期或是疾病、分裂素刺激下的蛋白质表达水平变化.然而，许多至关重要的生命进程不仅由蛋白质的相对丰度控制，更重要的是被那些时空特异分布的可逆翻译后修饰控制的，揭示翻译后修饰发生规律是理解蛋白质复杂多样的生物功能的一个重要前提.由于翻译后修饰蛋白质在样本中含量低且动态范围广，其相关研究极具挑战性，亲和富集、多维分离等技术与生物质谱的结合为翻译后修饰蛋白质组学的发展提供了契机，目前，已进行规模化研究的蛋白质翻译后修饰主要有四大类，其中磷酸化和糖基化研究较多.本文针对大规模翻译后的修饰蛋白质的分析策略和技术路线,如蛋白质的磷酸化修饰, 糖基化修饰, 泛素化修饰,基于蛋白质氧化还原状态进行的氧化还原修饰和其它修饰像乙酰化、甲基化、脂基化修饰等进行了综述.

Technique Strategies of Modification-specific Proteomics Study

Most work of early proteomics was focused on the protein expression profile of cells on the different growth period or cells perturbed by mitogens or diseases. However, many vital processes are controlled not only by the relative abundance of proteins but also by reversible post-translational modifications with specific spatial and temporal distributions. Consequently,it is important for comprehending complex biological functions of proteins to reveal the post-translational modification occurrence. Because of the low stoichiometry and broad dynamic range, research on the post-translationally modified proteins is still a big technical challenge. Nevertheless,development and integration of different methods, such as affinity-based enrichment methods,multi-dimensional separation technologies and mass spectrometry give the promise to probe the post-translational proteomics in a large scale. Currently, there are several major kinds of large-scale post-translational modification study, of which phosphorylation and glycosylation are researched more frequently. This article reviews the strategies and techniques used in high-throughput investigation of post-translationally modified proteins in recent years.