Abstract：
The misuse of poisonous mushrooms containing amatoxins causes acute liver failure (ALF) in patients and is a cause of significant mortality. Although the toxic mechanisms of α-amanitin (α-AMA) and its interactions with RNA polymerase II (RNAP II) have been studied, α-AMA effector proteins that can interact with α-AMA in hepatocytes have not been systematically studied. Limited proteolysis-coupled mass spectrometry (LiP-MS) is an advanced technology that can quickly identify protein-ligand interactions based on global comparative proteomics. This study identified the α-AMA effector proteins found in human hepatocytes, following the detection of conformotypic peptides using LiP-MS coupled with tandem mass tag (TMT) technology. Proteins that are classified into protein processing in the endoplasmic reticulum and the ribosome during the KEGG pathway can be identified through affinity evaluation, according to α-AMA concentration-dependent LiP-MS and LiP-MS in hepatocytes derived from humans and mice, respectively. The possibility of interaction between α-AMA and proteins containing conformotypic peptides was evaluated through molecular docking studies. The results of this study suggest a novel path for α-AMA to induce hepatotoxicity through interactions with various proteins involved in protein synthesis, as well as with RNAP II.
摘要：
误用含有阿马毒素的毒蘑菇会导致患者急性肝功能衰竭（ALF），并导致大量死亡。尽管已经研究了α-amanitin（α-AMA）的毒性机制及其与RNA聚合酶II（RNAPII）的相互作用，尚未系统地研究可以与肝细胞中的α-AMA相互作用的α-AMA效应蛋白。有限蛋白水解耦合质谱（LiP-MS）是一种基于全球比较蛋白质组学的先进技术，可以快速识别蛋白质-配体相互作用。这项研究确定了在人肝细胞中发现的α-AMA效应蛋白，在使用与串联质量标签(TMT)技术偶联的LiP-MS检测构象肽之后。在KEGG途径中被分类为内质网和核糖体中的蛋白质加工的蛋白质可以通过亲和力评估来鉴定，根据α-AMA浓度依赖性的LiP-MS和LiP-MS在人和小鼠肝细胞中的表达，分别。通过分子对接研究评估了α-AMA与含有构象肽的蛋白质之间相互作用的可能性。这项研究的结果表明，α-AMA通过与参与蛋白质合成的各种蛋白质相互作用来诱导肝毒性的新路径。以及RNAPII。
