BeyondSNO: Signalling beyond protein S-nitrosylation - determining the roles of nitroxyl and hydroxylamine

BeyondSNO：蛋白质 S-亚硝基化之外的信号传导 - 确定硝酰基和羟胺的作用

基本信息

批准号：
EP/Y027698/1
负责人：
Philip Eaton
金额：
$ 269.79万
依托单位：
Queen Mary University of London
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2024
资助国家：
英国
起止时间：
2024 至无数据
项目状态：
未结题

来源：
https://gtr.ukri.org/projects?ref=EP%2FY027698%2F1
关键词：
BeyondSNO Signalling beyond protein nitrosylation

项目摘要

Nitroxyl (HNO) is formed endogenously in cells, with drugs that release it in clinical development for heart failure. Despite this, it remains relatively unclear how this chemical relative of nitric oxide (NO) exerts its biological and therapeutic actions - an important issue we will address here. We will define the protein cysteines that nitroxyl post-translationally modifies to modulate cardiovascular function using redox proteomics. An underappreciated and yet undefined consequence of HNO modifying protein thiols is formation of hydroxylamine (NH2OH), which removes modifications such as S-palmitoylation from proteins. Consequently, we will also identify the proteins in heart and arteries that are removed by hydroxylamine formed when HNO is present. Identifying the target proteins modified by HNO and hydroxylamine will enable the cellular consequences of nitroxyl-driven signalling to be determined by making and functionally characterising 'redox dead' mutant proteins with the target cysteine mutated. Generation of novel 'redox dead' mice that lack these specific regulatory cysteines will allow the role of HNO and hydroxylamine signaling in cardiovascular health and disease to be established in vivo. We will leverage our findings therapeutically by identifying thiol reactive drugs, from our electrophilic compounds library, that target and alter the activity of the proteins modified by HNO or hydroxylamine. Such drugs are likely to have less side effects than nitroxyl donors that cause widespread off-target oxidations in addition to those that mediate their therapeutic actions. In contrast, the electrophilic drugs we identify will selectively modify the protein cysteine residue that mediates the therapeutic actions of HNO. The 'redox dead' mice we will generate will also be invaluable for drug discovery, as they will be resistant to the electrophilic drug because they lack the critical cysteine, providing robust therapeutic target validation.

硝酰基 (HNO) 是细胞内源性形成的，在治疗心力衰竭的临床开发中，药物会释放它。尽管如此，一氧化氮 (NO) 的这种化学相关物如何发挥其生物和治疗作用仍然相对不清楚——这是我们将在这里讨论的一个重要问题。我们将使用氧化还原蛋白质组学来定义硝酰基翻译后修饰以调节心血管功能的蛋白质半胱氨酸。 HNO 修饰蛋白质硫醇的一个未被充分认识且尚未明确的后果是形成羟胺 (NH2OH)，它会消除蛋白质中的 S-棕榈酰化等修饰。因此，我们还将鉴定心脏和动脉中的蛋白质，这些蛋白质会被硝酸存在时形成的羟胺去除。鉴定经 HNO 和羟胺修饰的目标蛋白将能够通过制造和功能表征具有目标半胱氨酸突变的“氧化还原死亡”突变蛋白来确定硝酰基驱动信号传导的细胞后果。缺乏这些特定调节半胱氨酸的新型“氧化还原死亡”小鼠的产生将允许在体内建立硝酸和羟胺信号传导在心血管健康和疾病中的作用。我们将通过从我们的亲电化合物库中鉴定硫醇反应药物来治疗我们的发现，这些药物靶向并改变经硝酸或羟胺修饰的蛋白质的活性。与硝酰基供体相比，此类药物的副作用可能更小，硝酰基供体除了介导其治疗作用外，还会引起广泛的脱靶氧化。相比之下，我们发现的亲电药物将选择性地修饰介导 HNO 治疗作用的蛋白质半胱氨酸残基。我们将产生的“氧化还原死亡”小鼠对于药物发现也将具有无价的价值，因为它们缺乏关键的半胱氨酸，因此会对亲电子药物产生抗药性，从而提供强大的治疗靶点验证。