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dc.contributor.authorKoen, Yakov M.
dc.contributor.authorSarma, Diganta
dc.contributor.authorWilliams, Todd D.
dc.contributor.authorGaleva, Nadezhda A.
dc.contributor.authorObach, R. Scott
dc.contributor.authorHanzlik, Robert P.
dc.date.accessioned2017-05-08T16:08:43Z
dc.date.available2017-05-08T16:08:43Z
dc.date.issued2012-05-21
dc.identifier.citationKoen, Y. M., Sarma, D., Williams, T. D., Galeva, N. A., Obach, R. S., & Hanzlik, R. P. (2012). Identification of Protein Targets of Reactive Metabolites of Tienilic Acid in Human Hepatocytes. Chemical Research in Toxicology, 25(5), 1145–1154. http://doi.org/10.1021/tx300103jen_US
dc.identifier.urihttp://hdl.handle.net/1808/23999
dc.descriptionThis document is the Accepted Manuscript version of a Published Work that appeared in final form in Chemical Research in Toxicology, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see http://pubs.acs.org/doi/abs/10.1021/tx300103jen_US
dc.description.abstractTienilic acid (TA) is a uricosuric diuretic that was withdrawn from the market only months after its introduction because of reports of serious incidents of drug-induced liver injury including some fatalities.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 [14C]-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 non-redundant 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 2–4 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 human 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.en_US
dc.publisherACSen_US
dc.rights© 2012 American Chemical Societyen_US
dc.titleIdentification of Protein Targets of Reactive Metabolites of Tienilic Acid in Human Hepatocytesen_US
dc.typeArticleen_US
kusw.kuauthorKoen, Yakov M.
kusw.kuauthorSarma, Diganta
kusw.kuauthorHanzlik, Robert P.
kusw.kuauthorWilliams, Todd D.
kusw.kuauthorGaleva, Nadezhda A.
kusw.kudepartmentMedicinal Chemistryen_US
kusw.kudepartmentMass Spectrometry Laboratoryen_US
kusw.oanotesPer SherpaRomeo on 05/08/2017: Author's Pre-print: grey tick subject to Restrictions below, author can archive pre-print (ie pre-refereeing) Restrictions: Must obtain written permission from Editor Must not violate ACS ethical Guidelines Author's Post-print: grey tick subject to Restrictions below, author can archive post-print (ie final draft post-refereeing) Restrictions: If mandated by funding agency or employer/ institution If mandated to deposit before 12 months, must obtain waiver from Institution/Funding agency or use AuthorChoice 12 months embargo Publisher's Version/PDF: cross author cannot archive publisher's version/PDF General Conditions: On author's personal website, pre-print servers, institutional website, institutional repositories or subject repositories Non-Commercial Must be accompanied by set statement (see policy) Must link to publisher version Publisher's version/PDF cannot be useden_US
dc.identifier.doi10.1021/tx300103jen_US
kusw.oaversionScholarly/refereed, author accepted manuscripten_US
kusw.oapolicyThis item meets KU Open Access policy criteria.en_US
dc.identifier.pmidPMC3358484en_US
dc.rights.accessrightsopenAccess


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