Sinclair Wilson R, Shrimp Jonathan H, Zengeya Thomas T, Kulkarni Rhushikesh A, Garlick Julie M, Luecke Hans, Worth Andrew J, Blair Ian A, Snyder Nathaniel W, Meier Jordan L
Chemical Biology Laboratory , Center for Cancer Research , National Cancer Institute , National Institutes of Health , Frederick , MD 21702 , USA . Email:
National Institute of Diabetes and Digestive and Kidney Diseases , National Institutes of Health , Bethesda , MD 20817 , USA.
Chem Sci. 2017 Dec 8;9(5):1236-1241. doi: 10.1039/c7sc00247e. eCollection 2018 Feb 7.
Short chain fatty acids (SCFAs) play a central role in health and disease. One function of these signaling molecules is to serve as precursors for short chain fatty acylation, a class of metabolically-derived posttranslational modifications (PTMs) that are established by lysine acetyltransferases (KATs) and lysine deacetylases (KDACs). this mechanism, short chain fatty acylation serves as an integrated reporter of metabolism as well as KAT and KDAC activity, and has the potential to illuminate the role of these processes in disease. However, few methods to study short chain fatty acylation exist. Here we report a bioorthogonal pro-metabolite strategy for profiling short chain fatty acylation in living cells. Inspired by the dietary component tributyrin, we synthesized a panel of ester-caged bioorthogonal short chain fatty acids. Cellular evaluation of these agents led to the discovery of an azido-ester that is metabolized to its cognate acyl-coenzyme A (CoA) and affords robust protein labeling profiles. We comprehensively characterize the metabolic dependence, toxicity, and histone deacetylase (HDAC) inhibitor sensitivity of these bioorthogonal pro-metabolites, and apply an optimized probe to identify novel candidate protein targets of short chain fatty acids in cells. Our studies showcase the utility of bioorthogonal pro-metabolites for unbiased profiling of cellular protein acylation, and suggest new approaches for studying the signaling functions of SCFAs in differentiation and disease.
短链脂肪酸（SCFAs）在健康和疾病中起着核心作用。这些信号分子的一个功能是作为短链脂肪酰化的前体，短链脂肪酰化是一类由赖氨酸乙酰转移酶（KATs）和赖氨酸脱乙酰酶（KDACs）建立的代谢衍生的翻译后修饰（PTMs）。通过这种机制，短链脂肪酰化作为代谢以及KAT和KDAC活性的综合报告因子，并且有可能阐明这些过程在疾病中的作用。然而，研究短链脂肪酰化的方法很少。在这里，我们报告了一种用于分析活细胞中短链脂肪酰化的生物正交前体代谢物策略。受膳食成分三丁酸甘油酯的启发，我们合成了一组酯笼蔽的生物正交短链脂肪酸。对这些试剂的细胞评估导致发现了一种叠氮基酯，它被代谢为其同源酰基辅酶A（CoA）并提供强大的蛋白质标记谱。我们全面表征了这些生物正交前体代谢物的代谢依赖性、毒性和组蛋白脱乙酰酶（HDAC）抑制剂敏感性，并应用优化的探针来鉴定细胞中短链脂肪酸的新型候选蛋白质靶点。我们的研究展示了生物正交前体代谢物在细胞蛋白质酰化无偏分析中的实用性，并提出了研究SCFAs在分化和疾病中的信号功能的新方法。
