UDP-glucosyltransferase activity toward exogenous substrates in Drosophila melanogaster.
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).
Real
M D
MD
Departamento de Genética, Universidad de Valencia, Spain.
Ferré
J
J
Chapa
F J
FJ
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004331:Drosophila melanogaster / Q000201:enzymology*; D005964:Glucosyltransferases / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D009284:Naphthols / Q000378:metabolism; D009596:Nitrophenols / Q000378:metabolism; D013379:Substrate Specificity; D014530:Uridine Diphosphate / Q000378:metabolism*
D009284:Naphthols; D009596:Nitrophenols; C029350:1-naphthol; D014530:Uridine Diphosphate; D005964:Glucosyltransferases; C028405:2-naphthol; C024836:4-nitrophenol