硫化氢介导的S-巯基化修饰及其化学检测技术

Hydrogen Sulfide Mediates Sulfhydration and Its ChemicalDetection Techniques

硫化氢(hydrogen sulfide, H_(2)S)是继一氧化氮和一氧化碳之后的第3种内源性气体信号分子,通过影响细胞信号通路调节机体各个系统,具有广泛的生理和病理作用。近年的研究发现,H2S的作用主要通过对靶蛋白进行S-巯基化修饰,改变其结构,影响其活性、稳定性以及蛋白质之间互相作用,进而调控细胞内信号通路及相关生物学过程。本文基于国内外对S-巯基化修饰的研究,主要综述了:S-巯基化修饰的研究现状,详细总结了目前已揭示S-巯基化修饰及其具体半胱氨酸(cysteine, Cys)位点的研究,并进一步总结了S-巯基化修饰与S-亚硝基修饰之间的关系;S-巯基化修饰的反应类型,3种化学检测(马来酰亚胺法、改良生物素转换法和标记转换法)方法的检测原理和特点,通过S-巯基化修饰检测的条件和应用范围,将这3种方法进行对比,并对各方法在检测过程中应注意的事项进行了讨论。根据作者在S-巯基化修饰研究方面积累的经验,重点阐述马来酰亚胺法检测过程中,细胞和组织样品的制备及其较详细的实验步骤。此外,本文还讨论了当前S-巯基化修饰检测方法存在的不足及该领域未来亟待解决的问题,为S-巯基化修饰的相关研究提供参考。

Hydrogen sulfide(H_(2)S)is the third endogenous gasotransmitter,following nitric oxide and carbon monoxide,which regulates various physiological and pathological processes by influencing cellular signaling pathways.In recent years,research has found that the effects of H 2S are mainly mediated through sulfhydration of target proteins,which can alter their structure,affect their activity,stability and protein-protein interactions,thus regulating cellular signaling pathways and related biological processes.This article reviews the current research of sulfhydration and provides a detailed summary of the known sulfhydration modifications and their specific cysteine(Cys)sites that have been identified.This article further discusses the relationship between sulfhydration and nitrosylation.It also covers the types of reactions involved in sulfhydration modification and describes the principles and characteristics of three chemical detection methods:the maleimide assay,the modified biotin switch assay,and the tag-switch assay.By comparing the conditions and applications of sulfhydration detection using these three methods,this article further summarizes the important procedures during the detection process.Based on our experience in sulfhydration research,this article focuses mainly on the cell and tissue sample preparation with detailed experimental procedures for the maleimide assay.Additionally,it discusses the limitations of current sulfhydration detection methods and the unresolved issues in this field,which may provide references for further research in sulfhydration.