Document Detail


Analysis of naphthalene adduct binding sites in model proteins by tandem mass spectrometry.
MedLine Citation:
PMID:  22659010     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The electrophilic metabolites of the polyaromatic hydrocarbon naphthalene have been shown to bind covalently to proteins and covalent adduct formation correlates with the cytotoxic effects of the chemical in the respiratory system. Although 1,2-naphthalene epoxide, naphthalene diol epoxide, 1,2-naphthoquinone, and 1,4-napthoquinone have been identified as reactive metabolites of interest, the role of each metabolite in total covalent protein adduction and subsequent cytotoxicity remains to be established. To better understand the target residues associated with the reaction of these metabolites with proteins, mass spectrometry was used to identify adducted residues following (1) incubation of metabolites with actin and protein disulfide isomerase (PDI), and (2) activation of naphthalene in microsomal incubations containing supplemental actin or PDI. All four reactive metabolites bound to Cys, Lys or His residues in actin and PDI. Cys₁₇ of actin was the only residue adducted by all metabolites; there was substantial metabolite selectivity for the majority of adducted residues. Modifications of actin and PDI, following microsomal incubations containing ¹⁴C-naphthalene, were detected readily by 2D gel electrophoresis and phosphor imaging. However, target modifications on tryptic peptides from these isolated proteins could not be readily detected by MALDI/TOF/TOF and only three modified peptides were detected using high resolution-selective ion monitoring (HR-SIM). All the reactive metabolites investigated have the potential to modify several residues in a single protein, but even in tissues with very high rates of naphthalene activation, the extent of modification was too low to allow unambiguous identification of a significant number of modified residues in the isolated proteins.
Authors:
Nathalie T Pham; William T Jewell; Dexter Morin; Alan R Buckpitt
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2012-05-31
Journal Detail:
Title:  Chemico-biological interactions     Volume:  199     ISSN:  1872-7786     ISO Abbreviation:  Chem. Biol. Interact.     Publication Date:  2012 Aug 
Date Detail:
Created Date:  2012-09-03     Completed Date:  2012-11-06     Revised Date:  2013-09-03    
Medline Journal Info:
Nlm Unique ID:  0227276     Medline TA:  Chem Biol Interact     Country:  Ireland    
Other Details:
Languages:  eng     Pagination:  120-8     Citation Subset:  IM    
Copyright Information:
Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.
Affiliation:
Department of Molecular Biosciences, School of Veterinary Medicine, University of California, Davis, CA 95616, USA. natpham@ucdavis.edu
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MeSH Terms
Descriptor/Qualifier:
Actins / chemistry,  metabolism*
Amino Acid Sequence
Animals
Binding Sites
Male
Mice
Microsomes, Liver / metabolism*
Molecular Sequence Data
Naphthalenes / chemistry,  metabolism*
Protein Binding
Protein Disulfide-Isomerases / chemistry,  metabolism*
Rats
Rats, Sprague-Dawley
Tandem Mass Spectrometry
Grant Support
ID/Acronym/Agency:
ES 04311/ES/NIEHS NIH HHS; ES 04699/ES/NIEHS NIH HHS; P42 ES004699/ES/NIEHS NIH HHS; R01 ES004311/ES/NIEHS NIH HHS
Chemical
Reg. No./Substance:
0/Actins; 0/Naphthalenes; 2166IN72UN/naphthalene; EC 5.3.4.1/Protein Disulfide-Isomerases
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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