Small intestinal UDP-glucuronosyltransferase sheUGT1A07: partial purification and cDNA cloning from sheep small intestine.
A phenol UDPglucuronosyltransferase (UGT) was partially purified, and the cDNA encoding the isoform was cloned and sequenced from sheep small intestine. The purified preparation containing a one major band (57 kDa) and one minor band (50 kDa) revealed high activities toward xenobiotics such as 1-naphthol (1-NA), 4-nitrophenol, and 4-methylumbelliferone. The preparation, however, had only little activity toward 4-hydroxybiphenyl and no activity toward bilirubin, suggesting that the preparation contains UGT1 isoforms. The NH2-terminal amino acid sequence of the major band was determined to be Gly-Lys-Leu-Leu-Val-Val-Pro-Met-Asp-Gly-Ser. A full-length UGT cDNA was obtained by reverse transcription-polymerase chain reaction with the degenerated 5'-primer from the NH2-terminal amino acid sequence of the purified major one and rapid amplification of cDNA ends from sheep small intestine. The cloned cDNA named sheUGT1A07 by amino acid similarity has a NH2-terminus sequence identical to that of the purified major one. Another phenol UGT cDNA named sheUGT1A6 was also cloned from sheep liver. sheUGT1A6 was expressed mainly in the liver, whereas sheUGT1A07 mRNA was expressed almost only in the alimentary organs, suggesting that sheUGT1A6 plays a role as a general drug metabolizing UGT isoform in the liver and sheUGT1A07 plays important role in the xenobiotics glucuronidation in the sheep small intestine.
Kobayashi
T
T
School of Veterinary Medicine, Rakuno Gakuen University, 582-1 Bunkyoudai-Midorimachi, Hokkaido, Ebetsu, 069-8501, Japan.
Tatano
A
A
Yokota
H
H
Onaga
T
T
Watanabe
T
T
Yuasa
A
A
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis; D014453:Glucuronosyltransferase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D007413:Intestinal Mucosa / Q000201:enzymology; D007421:Intestine, Small / Q000201:enzymology*; D007527:Isoenzymes / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D017421:Sequence Analysis; D012689:Sequence Homology, Nucleic Acid; D012756:Sheep; D014018:Tissue Distribution; D015262:Xenobiotics
D018076:DNA, Complementary; D007527:Isoenzymes; D015262:Xenobiotics; D014453:Glucuronosyltransferase