组氨酸磷酸化修饰的鉴定方法及应用

Technique development and applications of histidine phosphoproteomics

蛋白质磷酸化是广受关注的翻译后修饰类型之一,组氨酸磷酸化作为一种非常见的磷酸化修饰,最早被发现在细菌和低等真核生物信号传导的级联反应中起关键作用。近年来研究显示,其在肿瘤发生发展过程中也可能扮演了重要角色。由于磷酸化组氨酸的化学不稳定性、低丰度、亚化学计量性质、缺乏特异性的富集试剂,导致研究手段缺乏,限制了人们对磷酸化组氨酸修饰底物蛋白质的认识。随着磷酸化组氨酸抗体的设计以及富集和质谱等鉴定方法的发展,更多的磷酸化组氨酸修饰底物被鉴定,从而加速了对磷酸化组氨酸生物学功能的认识。本文介绍了磷酸化组氨酸的化学性质、主要生物学功能,并综述了磷酸化修饰底物富集和鉴定技术等方面的最新进展,同时也要看到组氨酸磷酸化修饰组学研究仍然存在巨大的技术挑战。

Protein phosphorylation is one of the most concerned types of post-translational modifications. Histidine phosphorylation is an unusual phosphorylation modification that plays a key role in the cascade of signal transduction in bacteria and lower eukaryotes, and has been shown to play an important role in tumorigenesis recently. Due to the chemical instability, low abundance, substoichiometric and lack of specific enrichment reagents, it was lack of approaches for the detection of histidine phosphorylation. Thus the knowledge on the modified protein substrates is limited. Increasing number of phosphorylated histidine modified substrates has been identified with the optimization of phosphorylated histidine specific antibodies, enrichment methods and mass spectrometry approaches, accelerating the understanding of the biological function of phosphorylated histidine. Here we highlighted the chemical properties, main biological functions of phosphorylated histidine, and reviewed the methods advancement for enrichment and identification of phosphorylated modified substrates. At the same time, we are still facing huge challenges in this field.