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 language | English |
|---|---|
| Pages (from-to) | 754-758 |
| Number of pages | 5 |
| Journal | Biological and Pharmaceutical Bulletin |
| Volume | 16 |
| Issue number | 8 |
| DOIs | |
| Publication status | Published - Jan 1 1993 |
Fingerprint
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 journal › Article
}
TY - JOUR
T1 - Purification and Characterization of a Morphine UDP-Glucuronyltransferase Isoform from Untreated Rat Liver
AU - Ishii, Yuji
AU - Oguri, Kazuta
AU - Yoshimura, Hidetoshi
PY - 1993/1/1
Y1 - 1993/1/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0027451337&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0027451337&partnerID=8YFLogxK
U2 - 10.1248/bpb.16.754
DO - 10.1248/bpb.16.754
M3 - Article
VL - 16
SP - 754
EP - 758
JO - Biological and Pharmaceutical Bulletin
JF - Biological and Pharmaceutical Bulletin
SN - 0918-6158
IS - 8
ER -