Proteomic analysis of diaminochlorotriazine adducts in wistar rat pituitary glands and LbetaT2 rat pituitary cells
Document Type
Article
Abstract
Atrazine (ATRA) is the most commonly applied herbicide in the United States and is frequently detected in drinking water at significant levels. After oral exposure, ATRA metabolism yields diaminochlorotriazine (DACT), an electrophilic molecule that has been shown to form covalent protein adducts. This research was designed to identify ATRA-induced protein adducts formed in the pituitary gland of ATRA-exposed rats and in DACT-exposed LbetaT2 rat pituitary cells. Immunohistochemistry showed diffuse cytoplasmic and nuclear staining in both pituitary sections and LbetaT2 cells indicating the formation of DACT protein adducts. Protein targets from both rat pituitaries and LbetaT2 cell culture were identified following two-dimensional electrophoresis (2DE), immunodetection, and matrix-assisted laser desorption ionization-time of flight mass spectrometry analysis. Western blots from both exposed rats and LbetaT2 cells revealed over 30 DACT-modified spots that were not present in control animals. Protein spots were matched to concurrently run 2DE gels stained with Sypro Ruby, excised, and in-gel-digested with trypsin. Mass spectrometry analysis of digest peptides resulted in the identification of 19 spots and 8 unique proteins in the rats and 21 spots and 19 unique proteins in LbetaT2 cells. The identified proteins present in both sample types included proteasome activator complex subunit 1, ubiquitin carboxyl-terminal hydrolase isozyme L1, tropomyosin, ERp57, and RNA-binding proteins. Each of these proteins contains active-site or solvent-exposed cysteine residues, making them viable targets for covalent modification by DACT.
Medical Subject Headings
Animals; Atrazine (analogs & derivatives, metabolism, toxicity); Cell Line; Female; Herbicides (toxicity); Pituitary Gland (drug effects, metabolism); Proteins (metabolism); Proteomics; Rats; Rats, Wistar
Publication Date
4-1-2008
Publication Title
Chemical research in toxicology
ISSN
0893-228X
Volume
21
Issue
4
First Page
844
Last Page
51
PubMed ID
18370413
Digital Object Identifier (DOI)
10.1021/tx700386f
Recommended Citation
Dooley, G P.; Reardon, K F.; Prenni, J E.; Tjalkens, R B.; Legare, M E.; Foradori, C D.; Tessari, J E.; and Hanneman, W H., "Proteomic analysis of diaminochlorotriazine adducts in wistar rat pituitary glands and LbetaT2 rat pituitary cells" (2008). Translational Neuroscience. 2556.
https://scholar.barrowneuro.org/neurobiology/2556