State Key Laboratory of Chemical Oncogenomics, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, Shenzhen, 518055, P. R. China.
Chembiochem. 2022 Feb 16;23(4):e202100628. doi: 10.1002/cbic.202100628. Epub 2021 Dec 30.
Fatty acids play fundamental structural, metabolic, functional, and signaling roles in all biological systems. Altered fatty acid levels and metabolism have been associated with many pathological conditions. Chemical probes have greatly facilitated biological studies on fatty acids. Herein, we report the development and characterization of an alkynyl-functionalized long-chain fatty acid-based sulfonyl fluoride probe for covalent labelling, enrichment, and identification of fatty acid-associated proteins in living cells. Our quantitative chemical proteomics show that this sulfonyl fluoride probe targets diverse classes of fatty acid-associated proteins including many metabolic serine hydrolases that are known to be involved in fatty acid metabolism and modification. We further validate that the probe covalently modifies the catalytically or functionally essential serine or tyrosine residues of its target proteins and enables evaluation of their inhibitors. The sulfonyl fluoride-based chemical probe thus represents a new tool for profiling the expression and activity of fatty acid-associated proteins in living cells.
脂肪酸在所有生物系统中发挥着基本的结构、代谢、功能和信号作用。脂肪酸水平和代谢的改变与许多病理状况有关。化学探针极大地促进了脂肪酸的生物学研究。在此，我们报告了一种炔基功能化的长链脂肪酸磺酰氟探针的开发和表征，用于在活细胞中对脂肪酸相关蛋白进行共价标记、富集和鉴定。我们的定量化学蛋白质组学研究表明，这种磺酰氟探针靶向多种脂肪酸相关蛋白，包括许多代谢丝氨酸水解酶，这些酶已知参与脂肪酸代谢和修饰。我们进一步验证了该探针能够共价修饰其靶蛋白的催化或功能必需的丝氨酸或酪氨酸残基，并能够评估其抑制剂。因此，基于磺酰氟的化学探针为在活细胞中分析脂肪酸相关蛋白的表达和活性提供了一种新工具。
