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Changes in endoplasmic reticulum stress proteins and aldolase A in cells exposed to dopamine
Author(s) -
Dukes April A.,
Van Laar Victor S.,
Cascio Michael,
Hastings Teresa G.
Publication year - 2008
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.2008.05392.x
Subject(s) - endoplasmic reticulum , calreticulin , unfolded protein response , oxidative stress , biochemistry , calnexin , mitochondrion , proteome , western blot , proteomics , chemistry , microbiology and biotechnology , biology , gene
In Parkinson’s disease, oxidative stress is implicated in protein misfolding and aggregation, which may activate the unfolded protein response by the endoplasmic reticulum (ER). Dopamine (DA) can initiate oxidative stress via H 2 O 2 formation by DA metabolism and by oxidation into DA quinone. We have previously shown that DA quinone induces oxidative protein modification, mitochondrial dysfunction in vitro, and dopaminergic cell toxicity in vivo and in vitro . In this study, we used cysteine‐ and lysine‐reactive fluorescent dyes with 2D difference in‐gel electrophoresis, mass spectrometry, and peptide mass fingerprint analysis to identify proteins in PC12 cell mitochondrial‐enriched fractions that were altered in abundance following DA exposure (150 μM, 16 h). Quantitative changes in proteins labeled with fluorescent dyes indicated increases in a subset of proteins after DA exposure: calreticulin, ERp29, ERp99, Grp58, Grp78, Grp94 and Orp150 (149–260%), and decreased levels of aldolase A (39–42%). Changes in levels of several proteins detected by 2D difference in‐gel electrophoresis were confirmed by western blot. Using this unbiased proteomics approach, our findings demonstrated that in PC12 cells, DA exposure leads to a cellular response indicative of ER stress prior to the onset of cell death, providing a potential link between DA and the unfolded protein response in the pathogenesis of Parkinson’s disease.