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Research ArticleArticle

Evaluation of Expression and Glycosylation Status of UGT1A10 in Supersomes and Intestinal Epithelial Cells with a Novel Specific UGT1A10 Monoclonal Antibody

Shingo Oda, Yukiko Kato, Masahiko Hatakeyama, Atsushi Iwamura, Tatsuki Fukami, Toshiyuki Kume, Tsuyoshi Yokoi and Miki Nakajima
Drug Metabolism and Disposition September 2017, 45 (9) 1027-1034; DOI: https://doi.org/10.1124/dmd.117.075291
Shingo Oda
Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kakuma-machi, Kanazawa, Japan (S.O., Y.K., T.F., T.Y., M.N.); CLEA Japan, Fujinomiya, Japan (M.H.); DMPK Research Laboratories, Sohyaku Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, Toda, Japan (A.I.); and Discovery Technology Laboratories, Sohyaku Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, Toda, Japan (T.K.)
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Yukiko Kato
Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kakuma-machi, Kanazawa, Japan (S.O., Y.K., T.F., T.Y., M.N.); CLEA Japan, Fujinomiya, Japan (M.H.); DMPK Research Laboratories, Sohyaku Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, Toda, Japan (A.I.); and Discovery Technology Laboratories, Sohyaku Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, Toda, Japan (T.K.)
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Masahiko Hatakeyama
Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kakuma-machi, Kanazawa, Japan (S.O., Y.K., T.F., T.Y., M.N.); CLEA Japan, Fujinomiya, Japan (M.H.); DMPK Research Laboratories, Sohyaku Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, Toda, Japan (A.I.); and Discovery Technology Laboratories, Sohyaku Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, Toda, Japan (T.K.)
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Atsushi Iwamura
Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kakuma-machi, Kanazawa, Japan (S.O., Y.K., T.F., T.Y., M.N.); CLEA Japan, Fujinomiya, Japan (M.H.); DMPK Research Laboratories, Sohyaku Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, Toda, Japan (A.I.); and Discovery Technology Laboratories, Sohyaku Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, Toda, Japan (T.K.)
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Tatsuki Fukami
Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kakuma-machi, Kanazawa, Japan (S.O., Y.K., T.F., T.Y., M.N.); CLEA Japan, Fujinomiya, Japan (M.H.); DMPK Research Laboratories, Sohyaku Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, Toda, Japan (A.I.); and Discovery Technology Laboratories, Sohyaku Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, Toda, Japan (T.K.)
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Toshiyuki Kume
Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kakuma-machi, Kanazawa, Japan (S.O., Y.K., T.F., T.Y., M.N.); CLEA Japan, Fujinomiya, Japan (M.H.); DMPK Research Laboratories, Sohyaku Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, Toda, Japan (A.I.); and Discovery Technology Laboratories, Sohyaku Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, Toda, Japan (T.K.)
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Tsuyoshi Yokoi
Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kakuma-machi, Kanazawa, Japan (S.O., Y.K., T.F., T.Y., M.N.); CLEA Japan, Fujinomiya, Japan (M.H.); DMPK Research Laboratories, Sohyaku Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, Toda, Japan (A.I.); and Discovery Technology Laboratories, Sohyaku Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, Toda, Japan (T.K.)
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Miki Nakajima
Drug Metabolism and Toxicology, Faculty of Pharmaceutical Sciences, Kanazawa University, Kakuma-machi, Kanazawa, Japan (S.O., Y.K., T.F., T.Y., M.N.); CLEA Japan, Fujinomiya, Japan (M.H.); DMPK Research Laboratories, Sohyaku Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, Toda, Japan (A.I.); and Discovery Technology Laboratories, Sohyaku Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, Toda, Japan (T.K.)
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Abstract

UDP-Glucuronosyltransferases (UGTs) are major phase II drug-metabolizing enzymes. Each member of the UGT family exhibits a unique but occasionally overlapping substrate specificity and tissue-specific expression pattern. Earlier studies have reported that human UGT1A10 is expressed in the gastrointestinal tract at the mRNA level, but the evaluation at the protein level, especially tissue or cellular localization, has lagged behind because of the lack of a specific antibody. In this study, we prepared a monoclonal antibody to UGT1A10 to elucidate the tissue/cellular distribution and interindividual variability of UGT1A10 protein expression. Western blot analysis revealed that the prepared antibody does not cross-react with any other human UGTs. Using this specific antibody, we observed that UGT1A10 protein is expressed in the small intestine but not in the liver or kidney. Immunohistochemical analysis revealed the expression of UGT1A10 protein in epithelial cells of the crypts and villi of the duodenum. In the small intestine microsomes from six individuals, the UGT1A10 protein levels exhibited 16-fold variability. Dopamine 3- and 4-glucuronidation, which is mainly catalyzed by UGT1A10 and by other UGT isoforms marginally, exhibited 50- to 65-fold variability, and they were not correlated with the UGT1A10 protein levels. Interestingly, the enzymatic activities of recombinant UGT1A10 in insect cells that were normalized to the UGT1A10 protein level were markedly lower than those in pooled human small intestine microsomes. Thus, the UGT1A10 antibody we generated made it possible to investigate the tissue/cellular distribution and interindividual variability of UGT1A10 protein expression for understanding the pharmacological and toxicological role of UGT1A10.

Footnotes

    • Received January 31, 2017.
    • Accepted June 29, 2017.
  • ↵1 Current affiliation: Department of Drug Safety Sciences, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan.

  • https://doi.org/10.1124/dmd.117.075291.

  • Copyright © 2017 by The American Society for Pharmacology and Experimental Therapeutics
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Drug Metabolism and Disposition: 45 (9)
Drug Metabolism and Disposition
Vol. 45, Issue 9
1 Sep 2017
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Research ArticleArticle

Evaluation of UGT1A10 protein in human tissues

Shingo Oda, Yukiko Kato, Masahiko Hatakeyama, Atsushi Iwamura, Tatsuki Fukami, Toshiyuki Kume, Tsuyoshi Yokoi and Miki Nakajima
Drug Metabolism and Disposition September 1, 2017, 45 (9) 1027-1034; DOI: https://doi.org/10.1124/dmd.117.075291

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Research ArticleArticle

Evaluation of UGT1A10 protein in human tissues

Shingo Oda, Yukiko Kato, Masahiko Hatakeyama, Atsushi Iwamura, Tatsuki Fukami, Toshiyuki Kume, Tsuyoshi Yokoi and Miki Nakajima
Drug Metabolism and Disposition September 1, 2017, 45 (9) 1027-1034; DOI: https://doi.org/10.1124/dmd.117.075291
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