Production of Isoform-Selective Antipeptide Antibodies toward UGT1.1r and UGT2B1
Abstract
We document here in that two UDP-glucuronosyltransferase (UGT) isoforms sensitive to phenobarbital are involved in morphine glucuronidation in Wistar and Sprague-Dawley rats. The hepatic microsomal morphine UGT activity in untreated Gunn rats was significantly less than that of untreated Wistar rats. Although the morphine UGT activity in the liver of Gunn rats was increased by phenobarbital (PB) treatment, this was significantly less than that in the liver of PB-treated Wistar rats. UGT1.1r is an isoform of morphine UGT in rat, and UGT2B1 is also considered an isoform of morphine UGT, because UGT2B1 (stably expressed in V79 cells) exhibited morphine UGT activity. We prepared specific antipeptide antibodies against UGT1.1r and UGT2B1. Using isoform-specific antipeptide antibodies, both UGT1.1r and UGT2B1 in Wistar and Sprague-Dawley rats were inducible by PB treatment. However, UGT1.1r is not present in the liver from Gunn rats. This study is the first demonstration that protein levels of two morphine UGT isoforms, UGT1.1r and UGT2B1, in the liver of Wistar and Sprague-Dawley rats are inducible by PB treatment.
Footnotes
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Send reprint requests to: Dr. Kazuta Oguri, Faculty of Pharmaceutical Sciences, Kyushu University-62, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-82, Japan.
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This work was supported in part by the Sasakawa Scientific Research Grant from the Japan Science Society. Presented at the 10th annual meeting of the Japanese Society for Xenobiotics, Ohmiya, Japan, November 1995 [Y. Ishii, A. Takami, K. Tsuruda, A. Kurogi, H. Yamada, and K. Oguri: Xenobiot. Metab. Dispos.10(Suppl.), 341 (abstr.) (1995)].
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↵2 A new nomenclature for the designation of UDP-glucuronosyltransferase (9) was used.
- Abbreviations used are::
- M-3-G
- morphine-3-glucuronide
- M-6-G
- morphine-6-glucuronide
- UGT
- UDP-glucuronosyltransferase
- PB
- phenobarbital
- UGT1.1r and UGT2B1
- gene products of UGT1.1r and UGT2B1
- KLH
- Keyhole Limpet Hemocyanin
- BSA
- bovine serum albumin
- CH-Sepharose 4B
- ω-carboxypentylamino Sepharose 4B
- CNBr
- cyanide bromide
- IgG
- immunoglobulin G
- PBS
- phosphate-buffered saline
- SDS-PAGE
- sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- PVDF
- polyvinylidene difluoride
- SDS
- sodium dodecyl sulfate
- TBS-Triton
- Tris-buffered saline:Triton X-100
- ECL
- enhanced chemiluminescence
- RT-PCR
- reverse transcriptase-polymerase chain reaction
- MC
- 3-methylcholanthrene
- Received July 10, 1996.
- Accepted November 13, 1996.
- The American Society for Pharmacology and Experimental Therapeutics
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Induction of Two UDP-Glucuronosyltransferase Isoforms Sensitive to Phenobarbital that are Involved in Morphine Glucuronidation
