斑马鱼蛋白酪氨酸硫酸化修饰的检测方法研究

Study on a detection method of protein tyrosine sulfation modification in zebrafish

蛋白酪氨酸硫酸化(protein tyrosine sulfation,PTS)是一种重要的翻译后修饰,调控生命活动中多种生理和病理过程,但由于PTS状态不稳定且目前缺乏有效的富集方法,因此在生物样品中难以进行有效地检测。本研究以模式动物斑马鱼(Danio rerio)为研究材料,利用Orbitrap Exploris 480高分辨质谱仪检测了斑马鱼胚胎发育早期总蛋白的酪氨酸硫酸化修饰水平,通过该方法共计检测到26种蛋白(包括膜蛋白、分泌蛋白、胞质蛋白和核蛋白等)存在潜在的29个酪氨酸硫酸化修饰位点。本研究建立了斑马鱼胚胎发育早期蛋白酪氨酸硫酸化修饰的检测方法,为探索生物体蛋白硫酸化修饰的作用机制奠定了技术基础。

Protein tyrosine sulfation(PTS)is an important post-translational modification that regulates a variety of physiological and pathological processes.However,PTS is unstable and lacks effective enrichment methods,which make it difficult to be detected in biological samples.In this study,we detected the tyrosine sulfation modification level of total proteins in developing zebrafish embryos by using high-resolution mass spectrometer,Orbitrap Exploris 480.A total of 26 proteins with tyrosine sulfation were detected,including membrane proteins,secreted proteins,cytoplasmic proteins and nucleoproteins.This study established a methodology in detecting protein tyrosine sulfation modification in embryonic zebrafish,which paved the way for evaluating the biological function of PTS.