Purification and Characterization of a Morphine UDP-Glucuronyltransferase Isoform from Untreated Rat Liver

Yuji Ishii, Kazuta Oguri, Hidetoshi Yoshimura

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

A morphine UDP-glucuronyltransferase was purified from liver microsomes of untreated Sprague-Dawley rats. A new gel, ω-(β-carboxypropionylamino)octyl Sepharose 4B, was prepared by coupling monomethylsuccinate with ω)-aminooctyl Sepharose 4B and this was used as an efficient tool for the separation of microsomal enzymes. Emulgen 911 solubilized microsomes were applied to a column packed with the gel and eluted at pH 7.4 while increasing KCl concentration in a stepwise manner. An isoform was further purified with UDP-hexanolamine Sepharose 4B gel. The purified UDP-glucuronyltransferase (morphine UGT of untreated rat, morphine UGTut) exhibited a molecular weight of 52000 on sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis and was capable of glucuronidating the 3-hydroxyl group of morphine. The isoform catalyzed to a small extent the glucuronidation of 4-hydroxybiphenyl; however, no glucuronidation activity towards androsterone, testosterone, bilirubin, 4-nitrophenol and the 6-hydroxyl group of morphine was observed. The difference in properties, compared with morphine UGT (molecular weight 56000) which was purified previously from phenobarbital-treated rats, is discussed.

Original languageEnglish
Pages (from-to)754-758
Number of pages5
JournalBiological and Pharmaceutical Bulletin
Volume16
Issue number8
DOIs
Publication statusPublished - Jan 1 1993

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Glucuronosyltransferase
Uridine Diphosphate
Sepharose
Morphine
Protein Isoforms
Gels
Hydroxyl Radical
Liver
Molecular Weight
Nonoxynol
Androsterone
Liver Microsomes
Phenobarbital
Microsomes
Bilirubin
Sodium Dodecyl Sulfate
Sprague Dawley Rats
Testosterone
Polyacrylamide Gel Electrophoresis
Enzymes

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Pharmaceutical Science

Cite this

Purification and Characterization of a Morphine UDP-Glucuronyltransferase Isoform from Untreated Rat Liver. / Ishii, Yuji; Oguri, Kazuta; Yoshimura, Hidetoshi.

In: Biological and Pharmaceutical Bulletin, Vol. 16, No. 8, 01.01.1993, p. 754-758.

Research output: Contribution to journalArticle

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