Wuya College of Innovation, Shenyang Pharmaceutical University, Shenyang, Liaoning 110016, P. R. China.
Department of Pharmacognosy and Utilization Key Laboratory of Northeast Plant Materials, School of Traditional Chinese Medicine, Shenyang Pharmaceutical University, Shenyang 110016, P. R. China.
ACS Chem Biol. 2022 Apr 15;17(4):873-882. doi: 10.1021/acschembio.1c00917. Epub 2022 Mar 30.
Human exposure to widespread furan-containing compounds (FCCs) has drawn much attention due to the high risk of their toxicities. Identifying adducted proteins resulting from the metabolic activation of FCCs is the core to learning the mechanism of FCCs' toxic action. We succeeded in establishing a metabolic activation-based chemoproteomic platform to map FCC-derived protein adducts in cultured primary hepatocytes treated with FCCs and to pinpoint the modification sites, using click chemistry but without alkynylation or azidation of FCCs to be investigated. The proposed platform was systematically verified by biomimetic synthesis, liver microsomal incubation, and primary hepatocyte culture. A mixture of furan, 2-methylfuran, and 2,5-dimethylfuran as model was tested by use of the established platform. A total of hepatic 171 lysine-based adducted proteins and 145 cysteine-based adducted proteins by the reactive metabolites of the three FCCs were enriched and identified (Byonic score ≥ 100). The target proteins were found to mainly participate in ATP synthesis. The technique was also successfully applied to furan-containing natural products. The established platform made it possible to profile covalently adducted proteins, because of potential exposure to a vast inventory of over two million of FCCs documented.
由于其毒性风险很高，人类接触广泛存在的呋喃类化合物（FCCs）引起了广泛关注。鉴定 FCC 代谢活化产生的加合物蛋白是了解 FCC 毒性作用机制的核心。我们成功地建立了一种基于代谢活化的化学蛋白质组学平台，用于在经 FCC 处理的培养原代肝细胞中绘制 FCC 衍生的蛋白质加合物，并确定修饰位点，使用点击化学，但无需对 FCC 进行炔基化或叠氮化以进行研究。该平台通过仿生合成、肝微粒体孵育和原代肝细胞培养进行了系统验证。使用所建立的平台对呋喃、2-甲基呋喃和 2,5-二甲基呋喃的混合物进行了测试。三种 FCCs 的反应性代谢物共富集和鉴定了 171 种肝赖氨酸加合物蛋白和 145 种半胱氨酸加合物蛋白（Byonic 评分≥100）。靶蛋白主要参与 ATP 合成。该技术也成功应用于含呋喃的天然产物。由于可能接触到记录的超过 200 万种 FCCs，因此建立的平台可以对共价加合物蛋白进行分析。
