title: Protein targets of acrylamide adduct formation in cultured rat dopaminergic cells.
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.
authors.affiliation: Canadian Rivers Institute and Department of Biology, University of New Brunswick, Saint John, New Brunswick E2L 4L5, Canada. richard.lopachin@einstein.yu.edu
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Martyniuk
authors.forename: April
authors.initials: A
authors.lastname: Feswick
authors.forename: Bin
authors.initials: B
authors.lastname: Fang
authors.forename: John M
authors.initials: JM
authors.lastname: Koomen
authors.forename: David S
authors.initials: DS
authors.lastname: Barber
authors.forename: Terrence
authors.initials: T
authors.lastname: Gavin
authors.forename: Richard M
authors.initials: RM
authors.lastname: Lopachin
mesh_terms: D020106:Acrylamide / Q000009:adverse effects* / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D002478:Cells, Cultured; D003545:Cysteine / Q000378:metabolism; D059290:Dopaminergic Neurons / Q000187:drug effects* / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000187:drug effects / Q000378:metabolism; D040901:Proteomics; D051381:Rats
chemical_list: D009419:Nerve Tissue Proteins; D020106:Acrylamide; D003545:Cysteine