Dynamics and regulation of cysteine S-sulfhydration in cellular functions

细胞功能中半胱氨酸S-硫酸化的动力学和调节

• 批准号: RGPIN-2016-04051
• 负责人: Yang, Guangdong
• 金额: $ 2.26万
• 依托单位: Laurentian University of Sudbury
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=669170
• 关键词: Dynamics; regulation; cysteine; sulfhydration; cellular

Hydrogen sulfide (H2S) is now considered as an endogenous gasotransmitter in a variety of cellular functions and physiological processes. Cystathionine gamma-lyase (CSE) is a major H2S-producing enzyme in mammalian cells. It is proposed that one of the signaling mechanisms of H2S is through the S-sulfhydration of reactive cysteine residues on target proteins by yielding a hydropersulfide moiety (-SSH), with the potential to confer a functional change. S-sulfhydration modulates diverse cellular pathways, including metabolic pathways, protein degradative processes, DNA damage repair, protein interaction and localization, and stress response pathways. This particular redox modification of cysteine by S-sulfhydration has been achieved using proteomics techniques by coupling a specific enrichment strategy (biotin-switch assay and/or tag-switch assay) with high-throughput mass spectrometry (MS) analysis. Our current understanding of the role of S-sulfhydration as a protein signaling modality is still in its infancy. Little is known about the nature of or even necessity for enzymatic mechanisms that may directly add or remove SH groups from cysteine thiols. *** My long-term goal is to elucidate the biological importance of H2S in order to improve our understanding of the regulation of a multitude of cellular functions, and to better train the next generation of researchers in this field. The short-term goal, through this research program, is to determine the biology and chemistry characteristic of cysteine S-sulfhydration with respect to H2S-regulated actin polymerization and cell migration/gap junction in endothelial cells. Mice deficient for CSE showed endothelial dysfunction, suggesting the importance of H2S in maintenance of endothelial integrity. My preliminary data showed that lack of H2S attenuates actin polymerization and endothelial cell migration. In addition, inhibition of thioredoxin 1 (Trx1) strengthened but knockdown of sulfhydryl oxidase expression inhibited actin S-sulfhydration. The objectives of this program are to 1) investigate the effects of the CSE/H2S system in actin polymerization and endothelial cell migration/gap junction, and explore H2S S-sulfhydration of actin-cofilin-profilin complex and the potential cysteine resides; 2), examine the regulation of Trx1 and sulfhydryl oxidase on actin S-sulfhydration/desulfhydation.******* The fact that a very large number of proteins are basally S-sulfhydrated suggests that S-sulfhydration is an important physiologic and pathologic signal. This program may lead to a breakthrough in the understanding of the biological relevance of H2S signaling in humans, and such an understanding will help reveal a novel physiologic posttranslational modification for proteins, which potentially influences a multitude of biological pathways.*********

现在，在各种细胞功能和生理过程中，硫化氢（H2S）现在被视为内源性气候递质。胱淀粉γ-乙酰酶（CSE）是哺乳动物细胞中主要产生的H2S酶。 有人提出，H2S的信号传导机制之一是通过产生氢食硫化物部分（-SSH）的反应性半胱氨酸残基的S-硫化，具有赋予功能变化的潜力。 S-硫化可以调节各种细胞途径，包括代谢途径，蛋白质降解过程，DNA损伤修复，蛋白质相互作用和定位以及应力反应途径。 通过使用高通量质谱法（MS）分析，使用蛋白质组学技术（Biotin-Switch分析和/或TAG-SWitch分析），使用蛋白质组学技术对半胱氨酸进行了这种特殊的氧化还原修饰。我们目前对S-磺化为蛋白质信号模式的作用的理解仍处于起步阶段。关于可以直接从半胱氨酸硫醇直接添加或去除SH组的酶机制的性质甚至是必要的知之甚少。 ***我的长期目标是阐明H2的生物学重要性，以提高我们对多种细胞功能的调节的理解，并更好地训练该领域的下一代研究人员。 通过该研究计划，短期目标是确定与H2S调节的肌动蛋白聚合和细胞迁移/间隙连接在内皮细胞中的生物学和化学特征。缺乏CSE的小鼠表现出内皮功能障碍，表明H2在维持内皮完整性中的重要性。 我的初步数据表明，缺乏H2会减弱肌动蛋白聚合和内皮细胞迁移。 另外，抑制硫氧还蛋白1（TRX1）增强但抑制了硫化氧化酶表达的抑制作用抑制了肌动蛋白S-硫化。 该程序的目标是1）研究CSE/H2S系统在肌动蛋白聚合和内皮细胞迁移/间隙连接中的影响，并探索肌动蛋白 - 核蛋白 - 易素蛋白复合物的H2S S-硫化和潜在的半胱氨酸含量； 2），检查Trx1和硫酰氧化酶对肌动蛋白S-硫化/脱硫酶的调节。******* *****，这一事实是，大量蛋白质基本上是S-硫化的，这表明S-硫化是一种重要的生理和病理学信号。该程序可能会导致对人类中H2S信号的生物学相关性的理解的突破，并且这种理解将有助于揭示蛋白质的新生理变化后修饰，这可能会影响多种生物学途径。*********************************************************************************************************