Abstract：
Exposure to organophosphorus pesticides (OP) can have chronic adverse effects that are independent of inhibition of acetylcholinesterase, the classic target for acute OP toxicity. In pure proteins, the organophosphorus pesticide chlorpyrifos oxon induces a cross-link between lysine and glutamate (or aspartate) with loss of water. Tubulin is particularly sensitive to OP-induced cross-linking. Our goal was to explore OP-induced cross-linking in a complex protein sample, MAP-rich tubulin from Sus scrofa and to test 8 OP for their capacity to promote isopeptide cross-linking. We treated 100 μg of MAP-rich tubulin with 100 μM chlorpyrifos, chlorpyrifos oxon, methamidophos, paraoxon, diazinon, diazoxon, monocrotophos, or dichlorvos. Each sample was separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and stained with Coomassie blue. Five gel slices (at about 30, 50, 150, and 300 kDa, and the top of the separating gel) were removed from the lanes for each of the eight OP samples and from untreated control lanes. These gel slices were subjected to in-gel trypsin digestion. MSMS fragmentation spectra of the tryptic peptides were examined for isopeptide cross-links. Sixteen spectra yielded convincing evidence for isopeptide cross-linked peptides. Ten were from the chlorpyrifos oxon reaction, 1 from dichlorvos, 1 from paraoxon, 1 from diazinon, and 3 from diazoxon. It was concluded that catalysis of protein cross-linking is a general property of organophosphorus pesticides and pesticide metabolites. Data are available via ProteomeXchange with identifier PXD034529.
摘要：
暴露于有机磷农药（OP）可能会产生慢性不良反应，这些不良反应与乙酰胆碱酯酶的抑制作用无关。急性OP毒性的经典目标。在纯蛋白质中，有机磷农药毒死rifoxon诱导赖氨酸和谷氨酸（或天冬氨酸）之间的交联，并失去水分。微管蛋白对OP诱导的交联特别敏感。我们的目标是探索复杂蛋白质样品中OP诱导的交联，来自Susscrofa的富含MAP的微管蛋白，并测试8OP促进异肽交联的能力。我们用100μM毒死rif处理了100μgMAP丰富的微管蛋白，毒死蜱oxon,甲胺磷,对氧磷,二嗪农,重氮,久效磷,或者敌敌畏.使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离每个样品并用考马斯蓝染色。五个凝胶切片（约30、50、150和300kDa，和分离凝胶的顶部）从八个OP样品中的每一个的泳道和未处理的对照泳道中移出。对这些凝胶切片进行凝胶内胰蛋白酶消化。检查胰蛋白酶肽的MSMS片段化谱的异肽交联。16个光谱为异肽交联肽提供了令人信服的证据。十种来自毒死蜱的oxon反应，1来自敌敌畏，1来自对氧磷，1来自二嗪农，和3来自二氮唑。结论是，蛋白质交联的催化作用是有机磷农药和农药代谢产物的一般性质。数据可通过ProteomeXchange获得，标识符为PXD034529。
