PXD033593-2

PXD033593 is an original dataset announced via ProteomeXchange.

Title	Dichlorvos-induced formation of isopeptide crosslinks between proteins in SH-SY5Y cells
Description	Chlorpyrifos oxon catalyzes the crosslinking of proteins via an isopeptide bond between lysine and glutamic acid or aspartic acid in studies with purified proteins. Our goal was to determine the crosslinking activity of the organophosphorus pesticide, dichlorvos. We developed a protocol for examining crosslinks in a complex protein mixture consisting of human SH-SY5Y cells exposed to 10 µM dichlorvos. The steps in our protocol included immunopurification of crosslinked peptides by binding to anti-isopeptide antibody 81D1C2, stringent washing of the immobilized complex, release of bound peptides from Protein G agarose with 50% acetonitrile 1% formic acid, liquid chromatography tandem mass spectrometry on an Orbitrap Fusion Lumos mass spectrometer, Protein Prospector searches of mass spectrometry data, and manual evaluation of candidate crosslinked dipeptides. We report a low quantity of dichlorvos-induced KD and KE crosslinked proteins in human SH-SY5Y cells exposed to dichlorvos. Cells not treated with dichlorvos had no detectable KD and KE crosslinked proteins. Proteins in the crosslink were low abundance proteins. In conclusion, we provide a protocol for testing complex protein mixtures for the presence of crosslinked proteins. Our protocol could be useful for testing the association between neurodegenerative disease and exposure to organophosphorus pesticides.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_09:07:17.990.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Lawrence Schopfer
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2022-05-04 09:16:00	ID requested
1	2023-10-24 11:11:54	announced
⏵ 2	2023-11-14 09:07:18	announced	2023-11-14: Updated project metadata.
3	2024-10-22 06:08:46	announced	2024-10-22: Updated project metadata.

Schopfer LM, Lockridge O, Dichlorvos-induced formation of isopeptide crosslinks between proteins in SH-SY5Y cells. Anal Biochem, 655():114844(2022) [pubmed]

submitter keyword: SHSY5Y human neuroblastoma cells, dichlorvos, mass spectrometry, manual evaluation, Protein Prospector, crosslink,isopeptide, crosslinks

Oksana Lockridge
contact affiliation	Eppley Institute University of Nebraska Medical Center
contact email	olockrid@unmc.edu
lab head
Lawrence Schopfer
contact affiliation	University of Nebraska Medical Center
contact email	lmschopf@unmc.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD033593
     1. Label: PRIDE project
     2. Name: Dichlorvos-induced formation of isopeptide crosslinks between proteins in SH-SY5Y cells