Purification and Properties of UDP-Glucuronyltransferase from Kidney Microsomes of β-Naphthoflavone-Treated Rat
| Autor: | Goshi Ishihara, Kazuhide Ohta, Akira Yuasa, Nobuyuki Ohgiya, Hiroshi Yokota |
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| Rok vydání: | 1989 |
| Předmět: |
Male
Molecular Sequence Data Glucuronidation Kidney Biochemistry Catalysis Sepharose Column chromatography beta-Naphthoflavone Affinity chromatography Microsomes Acetylglucosaminidase Animals Amino Acid Sequence Glucuronosyltransferase Molecular Biology Glycoproteins Benzoflavones Flavonoids chemistry.chemical_classification Gel electrophoresis Chromatography Rats Inbred Strains General Medicine Rats Amino acid Isoenzymes Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase chemistry Microsomes Liver Microsome Chromatography column |
| Zdroj: | The Journal of Biochemistry. 106:248-252 |
| ISSN: | 1756-2651 0021-924X |
| Popis: | Rat kidney microsomal UDP-glucuronyltransferase activities toward phenoic xenobiotics were enhanced about 4-5-fold by treatment of the animal with beta-naphthoflavone. The transferase activity toward serotonin, an endogenous substrate, was also enhanced about 7.5-fold. A form of UDP-glucuronyltransferase was purified from kidney microsomes of beta-naphthoflavone-treated rat by solubilization with sodium cholate and two steps of column chromatography, the first with DEAE-Toyopearl (fast flow rate liquid chromatography:FFLC) and the second with UDP-hexanolamine Sepharose 4B (affinity chromatography). These procedures gave about 39-fold purification and 11.5% yield of the transferase activity toward 1-naphthol. The preparation, tentatively termed "GT-2," was highly purified as judged from the single protein band (Mr 54,000) on sodium dodecylsulfate (SDS)-polyacrylamide slab gel electrophoresis. It catalyzed the glucuronidation of not only phenolic xenobiotics such as 1-naphthol, 4-nitrophenol, and 4-methylumbelliferone but also serotonin. From the result that apparent molecular weight of GT-2 was reduced to 50,000 by endo-beta-N-acetylglucosaminidase H (Endo H)-treatment, GT-2 was found to be a 50,000 Da polypeptide carrying "high mannose" type oligosaccharide chain(s). The NH2-terminal sequence of 20 residues of GT-2 was determined to be Asp-Lys-Leu-Leu-Val-Val-Pro-Gln-Asp-Gly-Ser-His-Trp-Leu-Ser-Met-Lys-Glu- Ile-Val . It was observed that there are two amino acids substitutions in the seven NH2-terminal residues in comparison with GT-1, which was purified from liver microsomes of 3-methylcholanthrene-treated rat. The NH2-terminal sequence of GT-2 was found to be homologous with the NH2-terminal sequence from the 26th to 46th amino acid residue of various UDP-glucuronyltransferase cloned by other investigators.(ABSTRACT TRUNCATED AT 250 WORDS) |
| Databáze: | OpenAIRE |
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