Purification and characterization of a morphine UDP-glucuronyltransferase isoform from untreated rat liver

Biol Pharm Bull. 1993 Aug;16(8):754-8. doi: 10.1248/bpb.16.754.


A morphine UDP-glucuronyltransferase was purified from liver microsomes of untreated Sprague-Dawley rats. A new gel, omega(beta-carboxypropionylamino)octyl Sepharose 4B, was prepared by coupling monomethylsuccinate with omega-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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chromatography, Ion Exchange
  • Electrophoresis, Polyacrylamide Gel
  • Glucuronosyltransferase / analysis*
  • Glucuronosyltransferase / isolation & purification
  • In Vitro Techniques
  • Ion Exchange Resins
  • Isoenzymes / analysis*
  • Isoenzymes / isolation & purification
  • Liver / enzymology*
  • Male
  • Microsomes, Liver / enzymology
  • Morphine / metabolism*
  • Proteins / analysis
  • Rats
  • Rats, Sprague-Dawley
  • Sepharose
  • Substrate Specificity


  • Ion Exchange Resins
  • Isoenzymes
  • Proteins
  • Morphine
  • Sepharose
  • Glucuronosyltransferase