Document Detail


Identification of protein targets of reactive metabolites of tienilic acid in human hepatocytes.
MedLine Citation:
PMID:  22462724     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Tienilic acid (TA) is a uricosuric diuretic that was withdrawn from the market only months after its introduction because of reports of serious incidents of drug-induced liver injury including some fatalities. Its hepatotoxicity is considered to be primarily immunoallergic in nature. Like other thiophene compounds, TA undergoes biotransformation to a S-oxide metabolite which then reacts covalently with cellular proteins. To identify protein targets of TA metabolites, we incubated [(14)C]-TA with human hepatocytes, separated cellular proteins by 2D gel electrophoresis, and analyzed proteins in 36 radioactive spots by tryptic digestion followed by LC-MS/MS. Thirty-one spots contained at least one identifiable protein. Sixteen spots contained only one of 14 nonredundant proteins which were thus considered to be targets of TA metabolites. Six of the 14 were also found in other radioactive spots that contained from 1 to 3 additional proteins. Eight of the 14 had not been reported to be targets for any reactive metabolite other than TA. The other 15 spots each contained from 2 to 4 identifiable proteins, many of which are known targets of other chemically reactive metabolites, but since adducted peptides were not observed, the identity of the adducted protein(s) in these spots is ambiguous. Interestingly, all the radioactive spots corresponded to proteins of low abundance, while many highly abundant proteins in the mixture showed no radioactivity. Furthermore, of approximately 16 previously reported protein targets of TA in rat liver ( Methogo, R., Dansette, P., and Klarskov, K. ( 2007 ) Int. J. Mass Spectrom. , 268 , 284 -295 ), only one (fumarylacetoacetase) is among the 14 targets identified in this work. One reason for this difference may be statistical, given that each study identified a small number of targets from among thousands present in hepatocytes. Another may be the species difference (i.e., rat vs human), and still another may be the method of detection of adducted proteins (i.e., Western blot vs C-14). Knowledge of human target proteins is very limited. Of more than 350 known protein targets of reactive metabolites, only 42 are known from humans, and only 21 of these are known to be targets for more than one chemical. Nevertheless, the demonstration that human target proteins can be identified using isolated hepatocytes in vitro should enable the question of species differences to be addressed more fully in the future.
Authors:
Yakov M Koen; Diganta Sarma; Todd D Williams; Nadezhda A Galeva; R Scott Obach; Robert P Hanzlik
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2012-04-10
Journal Detail:
Title:  Chemical research in toxicology     Volume:  25     ISSN:  1520-5010     ISO Abbreviation:  Chem. Res. Toxicol.     Publication Date:  2012 May 
Date Detail:
Created Date:  2012-05-21     Completed Date:  2012-09-10     Revised Date:  2014-09-16    
Medline Journal Info:
Nlm Unique ID:  8807448     Medline TA:  Chem Res Toxicol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1145-54     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Electrophoresis, Gel, Two-Dimensional
Electrophoresis, Polyacrylamide Gel
Hepatocytes / metabolism*
Humans
Proteins / chemistry,  metabolism*
Tandem Mass Spectrometry
Ticrynafen / chemistry,  metabolism*
Grant Support
ID/Acronym/Agency:
GM-21784/GM/NIGMS NIH HHS; R01 GM021784/GM/NIGMS NIH HHS; R01 GM021784-33/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Proteins; HC95205SY4/Ticrynafen
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