Protein targets of acrylamide adduct formation in cultured rat dopaminergic cells

Christopher J. Martyniuk, April Feswick, Bin Fang, John M. Koomen, David S. Barber, Terrence Gavin, Richard M. LoPachin

Research output: Contribution to journalArticle

24 Scopus citations

Abstract

Acrylamide (ACR) is an electrophilic unsaturated carbonyl derivative that produces neurotoxicity by forming irreversible Michael-type adducts with nucleophilic sulfhydryl thiolate groups on cysteine residues of neuronal proteins. Identifying specific proteins targeted by ACR can lead to a better mechanistic understanding of the corresponding neurotoxicity. Therefore, in the present study, the ACR-adducted proteome in exposed primary immortalized mesencephalic dopaminergic cells (N27) was determined using tandem mass spectrometry (LTQ-Orbitrap). N27 cells were characterized based on the presumed involvement of CNS dopaminergic damage in ACR neurotoxicity. Shotgun proteomics identified a total of 15,243 peptides in N27 cells of which 103 unique peptides exhibited ACR-adducted Cys groups. These peptides were derived from 100 individual proteins and therefore ~0.7% of the N27 cell proteome was adducted. Proteins that contained ACR adducts on multiple peptides included annexin A1 and pleckstrin homology domain-containing family M member 1. Sub-network enrichment analyses indicated that ACR-adducted proteins were involved in processes associated with neuron toxicity, diabetes, inflammation, nerve degeneration and atherosclerosis. These results provide detailed information regarding the ACR-adducted proteome in a dopaminergic cell line. The catalog of affected proteins indicates the molecular sites of ACR action and the respective roles of these proteins in cellular processes can offer insight into the corresponding neurotoxic mechanism.

Original languageEnglish (US)
Pages (from-to)279-287
Number of pages9
JournalToxicology Letters
Volume219
Issue number3
DOIs
StatePublished - May 14 2013

Keywords

  • Nerve terminal
  • Neurotoxicity
  • Protein adduct formation
  • Proteomic analysis
  • Toxic neuropathy

ASJC Scopus subject areas

  • Toxicology

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