A Metabolic Activation-Based Chemoproteomic Platform to Profile Adducted Proteins Derived from Furan-Containing Compounds
A Metabolic Activation-Based Chemoproteomic Platform to Profile Adducted Proteins Derived from Furan-Containing Compounds
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DOI：
10.1021/acschembio.1c00917
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发表时间：
2022-04-15
影响因子：
4
通讯作者：
Zheng, Jiang
中科院分区：
文献类型：
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作者：
Li, Wei;Hu, Zixia;Zheng, Jiang
含呋喃化合物（FCCs）广泛存在于人体中，其高毒性引起了人们的广泛关注。鉴定FCC代谢活化过程中的加合蛋白质有助于了解FCC的毒性作用机制。我们成功地建立了一个基于代谢活化的化学蛋白质组学平台，在培养的原代肝细胞中用FCC处理FCC衍生的蛋白加合物，并精确定位修饰位点，使用点击化学，但没有对FCC进行炔基化或叠氮化研究。通过仿生合成、肝微粒体孵育和原代肝细胞培养，系统地验证了所提出的平台。以呋喃、2-甲基呋喃和2，5-二甲基呋喃的混合物为模型，利用所建立的实验平台进行了实验。共富集并鉴定了171个赖氨酸加合蛋白和145个半胱氨酸加合蛋白（Byonic评分>= 100）。目的蛋白主要参与ATP的合成。该技术也成功地应用于含呋喃的天然产物。建立的平台使分析共价加合蛋白成为可能，因为潜在的暴露于超过200万的FCC记录的巨大库存。
Human exposure to widespread furan-containing compounds (FCCs) has drawn much attention due to the high riskof their toxicities. Identifying adducted proteins resulting from themetabolic activation of FCCsisthecoretolearningthemechanism of FCCs'toxic action. We succeeded in establishinga metabolic activation-based chemoproteomic platform to mapFCC-derived protein adducts in cultured primary hepatocytestreated with FCCs and to pinpoint the modification sites, usingclick chemistry but without alkynylation or azidation of FCCs tobe investigated. The proposed platform was systematically verifiedby biomimetic synthesis, liver microsomal incubation, and primaryhepatocyte culture. A mixture of furan, 2-methylfuran, and 2,5-dimethylfuran as model was tested by use of the establishedplatform. A total of hepatic 171 lysine-based adducted proteins and 145 cysteine-based adducted proteins by the reactive metabolitesof the three FCCs were enriched and identified (Byonic score >= 100). The target proteins were found to mainly participate in ATPsynthesis. The technique was also successfully applied to furan-containing natural products. The established platform made itpossible to profile covalently adducted proteins, because of potential exposure to a vast inventory of over two million of FCCs documented.