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PMID:1898728

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Citation

Ritter, JK, Crawford, JM and Owens, IS (1991) Cloning of two human liver bilirubin UDP-glucuronosyltransferase cDNAs with expression in COS-1 cells. J. Biol. Chem. 266:1043-7

Abstract

We report the isolation and characterization of two human liver cDNA clones, HUG-Br1 and HUG-Br2; each encodes a UDP-glucuronosyltransferase enzyme which glucuronidates bilirubin IX alpha to form both the IX alpha C8 and IX alpha C12 monoconjugates and a diconjugate. HUG-Br1 cDNA (2351 base pairs) and HUG-Br2 cDNA (2368 base pairs) encode proteins with 533 and 534 amino acid residues, respectively, with a typical membrane-insertion signal peptide, membrane-spanning domain, and 3 or 5 potential asparagine-linked glycosylation sites. At the nucleic acid and deduced amino acid sequence levels the two clones are 82% similar overall, 66% similar in the amino termini, and identical after codon 287, thus encoding proteins with the same carboxyl terminus. The mRNA encoding HUG-Br1 is of high abundance, and the one encoding HUG-Br2 is of low abundance; both are 2.6 kilobases in length. Both messages (2.6 kilobases) were present in the explanted liver of a Type I Crigler-Najjar patient, although the level for that of HUG-Br1 was reduced 4.5-fold. Northern blot analysis of poly(A)+ RNA isolated from the liver of an untreated and a phenobarbital-treated Erythrocebus patas monkey with 5'-specific probes for each clone indicated that the HUG-Br2-encoded message is induced two fold, but that for HUG-Br1 is not. These data indicate that bilirubin is glucuronidated by at least two different proteins, most likely present in very different amounts. These cDNAs which encode functional bilirubin UDP-glucuronosyltransferases will allow the isolation of an appropriate gene to develop a gene therapy model for patients which have the totally deficient trait.

Links

PubMed

Keywords

Amino Acid Sequence; Animals; Base Sequence; Bilirubin/genetics; Bilirubin/metabolism; Blotting, Northern; Cell Line; Cloning, Molecular; DNA/genetics; DNA/isolation & purification; Erythrocebus patas; Gene Expression; Glucuronosyltransferase/genetics; Humans; Isoenzymes/genetics; Liver/enzymology; Microsomes, Liver/enzymology; Molecular Sequence Data; Nucleic Acid Hybridization; RNA, Messenger/genetics; Transfection

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

HUMAN:UD11

enables

GO:0015020: glucuronosyltransferase activity

ECO:0005801: enzymatic activity assay evidence used in manual assertion

F

Seeded From UniProt

complete

HUMAN:UD14

enables

GO:0015020: glucuronosyltransferase activity

ECO:0005801: enzymatic activity assay evidence used in manual assertion

F

Seeded From UniProt

complete

See also

References

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