UDP-glucosyltransferase activity toward exogenous substrates in Drosophila melanogaster
✍ Scribed by M.D. Real; J. Ferré; F.J. Chapa
- Publisher
- Elsevier Science
- Year
- 1991
- Tongue
- English
- Weight
- 407 KB
- Volume
- 194
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
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).