Proteomics

Dataset Information

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Organophosphorus pesticides catalyze protein crosslinking


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 crosslink between lysine and glutamate (or aspartate) with loss of water. Tubulin is particularly sensitive to OP-induced crosslinking. Our goal was to explore OP-induced crosslinking in a complex protein sample, MAP-rich tubulin from Sus scrofa, and to test 8 OP for their capacity to catalyze isopeptide crosslinking. 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 on SDS PAGE 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 crosslinks. Sixteen spectra yielded convincing evidence for isopeptide crosslinked 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 crosslinking is a general property of organophosphorus pesticides and pesticide metabolites.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Sus Scrofa Domesticus (domestic Pig)

SUBMITTER: Lawrence Schopfer  

LAB HEAD: Oksana Lockridge

PROVIDER: PXD034529 | Pride | 2022-10-14

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
20190402_Larry_2A_CPO.pdf Pdf
20190402_Larry_2A_CPO.raw Raw
20190402_Larry_2B_CPO.pdf Pdf
20190402_Larry_2B_CPO.raw Raw
20190402_Larry_9A_Diclorvos.pdf Pdf
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Publications

Organophosphorus Pesticides Promote Protein Cross-Linking.

Schopfer Lawrence M LM   Onder Seda S   Lockridge Oksana O  

Chemical research in toxicology 20220901 9


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 <i>Su  ...[more]

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