Purification and partial characterization of rat liver bilirubin glucuronoside glucuronosyltransferase

Jayanta Roy-Chowdhury, N. R. Chowdhury, M. M. Bhargava, I. M. Arias

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Abstract

Bilirubin glucuronoside glucuronosyltransferase (EC 2.4.1.95) converts bilirubin monoglucuronide to bilirubin diglucuronide and is concentrated in plasma membrane-enriched fractions of rat liver homogenates. The enzyme was purified 2,000-fold to homogeneity from rat liver. The pI of the enzyme is 7.9 ± 0.2. The enzyme has a molecular weight of 160,000 and is an oligomer of 28,000 dalton subunits. K(m) for purified enzyme was 35 μM and V(max) was 2.2 μmol of bilirubin diglucuronide formed/min/mg of protein. Freshly biosynthesized bilirubin monoglucuronide was injected intravenously into homozygous Gunn rats which had bile duct cannulation. Gunn rats lack UDP-glucuronate glucuronyltransferase activity (EC 2.4.1.17), have normal bilirubin glucuronoside glucuronosyltransferase activity, cannot form bilirubin monoglucuronide in vitro or in vivo, and do not excrete bilirubin glucuronides after intravenous injection of unconjugated bilirubin. Within 1 h, approximately 75% of the injected conjugated bilirubin was recovered in bile, of which 20% consisted of bilirubin diglucuronide. These results indicate that bilirubin glucuronoside glucuronosyltransferase catalyzes conversion of bilirubin monoglucuronide to diglucuronide in vivo.

Original languageEnglish (US)
Pages (from-to)8336-8339
Number of pages4
JournalJournal of Biological Chemistry
Volume254
Issue number17
StatePublished - 1979

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bilirubin glucuronoside glucuronosyltransferase
Liver
Purification
Rats
Gunn Rats
Enzymes
Bilirubin
Glucuronosyltransferase
Glucuronic Acid
Uridine Diphosphate
Cell membranes
Bile Ducts
Oligomers
Bile
Intravenous Injections
Catheterization
Ducts
Molecular Weight
Molecular weight
Cell Membrane

ASJC Scopus subject areas

  • Biochemistry

Cite this

Purification and partial characterization of rat liver bilirubin glucuronoside glucuronosyltransferase. / Roy-Chowdhury, Jayanta; Chowdhury, N. R.; Bhargava, M. M.; Arias, I. M.

In: Journal of Biological Chemistry, Vol. 254, No. 17, 1979, p. 8336-8339.

Research output: Contribution to journalArticle

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