Purification of a phenobarbital-inducible UDP-glucuronosyltransferase isoform from dog liver which catalyzes morphine and testosterone glucuronidation

Kazuta Oguri, Akiko Kurogi, Kou Ichi Yamabe, Mitsuko Tanaka, Kunihiro Yoshisue, Yuji Ishii, Hidetoshi Yoshimura

研究成果: Contribution to journalArticle査読

26 被引用数 (Scopus)

抄録

A morphine UDP-glucuronosyltransferase (UGT) which could belong to the UGT2B subfamily was isolated from liver microsomes of a male beagle dog treated with phenobarbital. Glucuronidation toward morphine in the dog liver microsomes was increased threefold by the treatment. The microsomes were solubilized with Emulgen 911 and applied on a column of hemisuccinate derivative of Sepharose 4B column which has been developed in our laboratory. An isoform of UGT in the eluate was purified further by chromatofocusing and UDP-hexanolamine-affinity chromatography. A purified enzyme, UGT(DOG-PB), was homogeneous on sodium dodecyl sulfate polyacrylamide gel electrophoresis and two-dimensional electrophoresis and exhibited a subunit molecular weight of 50 kDa. This isoform showed activities toward the 3-hydroxyl group of morphine, 4-hydroxybiphenyl, 4-nitrophenol, 4-methylumbelliferone, and testosterone, but not toward chloramphenicol and the 6-hydroxyl group of morphine. The substrate specificity of UGT(DOG-PB) is similar to that of stably expressed UGT2B1 which is considered a phenobarbital-inducible morphine UGT in the rat except that UGT(DOG-PB) is capable of glucuronidating 4-nitrophenol but not chloramphenicol. The NH2-terminus until the 30th residue of UGT(DOG-PB) is highly homologous to UGT2B subfamily, and the NH2- terminal 15 residues of UGT(DOG-PB) are completely identical to those of UGT2B1, UGT2B8, and UGT2B15. This is the first report describing the UGT isoform of dog and the purification of morphine UGT which may belong to UGT2B subfamily.

本文言語英語
ページ(範囲)159-166
ページ数8
ジャーナルArchives of Biochemistry and Biophysics
325
2
DOI
出版ステータス出版済み - 1 15 1996

All Science Journal Classification (ASJC) codes

  • 生物理学
  • 生化学
  • 分子生物学

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