6533b7d0fe1ef96bd125a3ca
RESEARCH PRODUCT
UDP-glucosyltransferase activity toward exogenous substrates in Drosophila melanogaster.
F.j. ChapaM.d. RealJuan Ferrésubject
BiophysicsNaphtholsBiochemistryHigh-performance liquid chromatographyUridine DiphosphateSubstrate SpecificityAbsorbanceNitrophenolschemistry.chemical_compoundGlucosideDrosophilidaeAnimalsMolecular BiologyChromatography High Pressure Liquidchemistry.chemical_classificationChromatographybiologySubstrate (chemistry)Cell BiologyHydrogen-Ion Concentrationbiology.organism_classificationFluorescenceEnzymeDrosophila melanogasterchemistryBiochemistryGlucosyltransferasesDrosophila melanogasterdescription
To investigate the capacity of Drosophila extracts to glucosylate exogenous substrates we have developed a fast and sensitive method for the detection of UDP-glucosyltransferase activity using 4-nitrophenol, 1-naphthol, or 2-naphthol as substrates. High-performance liquid chromatography was used to separate and quantitate the reaction products, allowing detection of activities that produced as little as 1 pmol of 2-naphthol glucoside (fluorescence detection) or 16 pmol of 4-nitrophenol glucoside (absorbance detection). Optimal activity was found at 43 degrees C and alkaline pH. The affinity of the Drosophila enzyme was 250-fold higher for 1-naphthol or 2-naphthol (Km approximately 4 microM) than for 4-nitrophenol and UDP-glucose (Km approximately 1 mM).
| year | journal | country | edition | language |
|---|---|---|---|---|
| 1991-05-01 | Analytical biochemistry |