A fluorescent assay amenable to measuring production of beta-D-glucuronides produced from recombinant UDP-glycosyl transferase enzymes

Drug Metab Dispos. 1999 May;27(5):555-7.

Abstract

Beta-glucuronidase cleavage of 4-methylumbelliferyl beta-D-glucuronide generates the highly fluorescent compound, 4-methylumbelliferone. We show that other beta-D-glucuronide compounds act as competitors in this assay. The 4-methylumbelliferyl beta-D-glucuronide cleavage assay can easily be adapted to high throughput formats to detect the presence of beta-D glucuronides generated using recombinant glycosyl transferase preparations.

MeSH terms

  • Binding, Competitive
  • Fluorescence
  • Glucuronates / analysis*
  • Glucuronates / metabolism*
  • Glucuronidase / genetics
  • Glucuronidase / metabolism
  • Glucuronosyltransferase / genetics
  • Glucuronosyltransferase / metabolism*
  • Hymecromone / analogs & derivatives
  • Hymecromone / metabolism
  • Kinetics
  • Recombinant Proteins / metabolism*

Substances

  • Glucuronates
  • Recombinant Proteins
  • Hymecromone
  • 4-methylumbelliferyl glucuronide
  • UDP-glucuronosyltransferase, UGT1A6
  • Glucuronosyltransferase
  • Glucuronidase