Instrument supplement: LCMS for MIRA-funded chemical biology research program

项目摘要 蛋白质脂化是一种影响亚细胞的动态翻译后修饰(PTM) 蛋白质的运输、辅因子结合亲和力和膜定位，进而 影响下游信号级联，特别是环状的S-酰化和-脱酰化 蛋白质和特定的胱氨酸残基正在成为循环中血脂水平和 对基本生物过程的调节，包括那些涉及细胞生长的过程， 代谢和神经健康，然而，对这种PTM的深入研究在技术上证明了 困难，在很大程度上是因为缺乏选择性的、有效的酶化学抑制剂 催化其安装和移除，拟议的研究计划旨在 产生新的化学技术，即小分子探针和抑制剂 说明调节蛋白S-酰化在正常和非正常人群中的作用 在古生理学背景下，这些目标将通过两种互补的化学物质来实现 和细胞生物学研究领域，其中一个领域将涉及测量、操作和 确定S-酰化的“作者”的靶标 将使用新开发和优化的筛选和选择性分析来识别抑制剂 与合理和计算设计的分子相结合的分析，以及 然后将使用假定的抑制剂、已验证的抑制剂和基于蛋白质组学的方法的库 确定活细胞中DHHC的特定蛋白质靶点，以更准确地描述其 参与各种疾病状态，第二个研究领域将利用我们最近验证的 用化学工具研究S-酰化反应中的“擦除器”的生物功能 重视这些酶在细胞氧化还原、动态平衡和代谢中的作用 疾病，这一多学科研究计划的预期结果是双重的：产生 用于研究这一重要的PTM的化学工具和分析方法的集合，并描述其 在正常和疾病状态下的生物学作用和影响，

Project Summary Protein lipidation is a dynamic post-translational modification (PTM) that affects subcellular trafficking, co-factor binding affinity, and membrane localization of proteins, which in turn influence downstream signaling cascades, In particular, cyclic S-acylation and -deacylation of proteins al specific cysleine residues is emerging as a key link between circulating lipid levels and the regulation of essential biological processes, including those involved in cellular growth, metabolism, and neurological health, In-depth study of this PTM, however, has proven technically difficult, in large part due to the paucity of selective, effective chemical inhibitors for the enzymes that catalyze its installation and removal, The proposed research program is designed to generate novel chemical technologies, namely small molecule probes and inhibitors, in the service of illuminating the involvement of regulated protein S-acylation in both normal and palhophysiological contexts, These goals will be realized through two complementary chemical and cellular biology research areas, One area will involve measuring, manipulating, and determining the targets of the "writers" of S-acylation, DHHCs, To do so, pan-active DHHC inhibitors will be identified using newly developed and optimized screening and selectivity profiling assays in combination with rationally and computationally designed molecules, as well as a library of putative inhibitors, Validated inhibitors and proteomics-based methods will then be used to identify the specific protein targets of DHHCs in live cells to more precisely describe their involvement in various disease states, The second research area will utilize our recently validated chemical tools to study the biological function of the "erasers" of S-acylation, APTs, with particular emphasis on the involvement of these enzymes in cellular redox homeostasis and metabolic disease, The expected outcome of this multidisciplinary research program is two-fold: generating a collection of chemical tools and assays for the study of this important PTM, and describing its biological function and influence in normal and disease states,