UDP-GLUCOSYLTRANSFERASE ACTIVITY TOWARD EXOGENOUS SUBSTRATES IN DROSOPHILA-MELANOGASTER
UDP-GLUCOSYLTRANSFERASE ACTIVITY TOWARD EXOGENOUS SUBSTRATES IN DROSOPHILA-MELANOGASTER
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DOI：
10.1016/0003-2697(91)90239-p
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发表时间：
1991-05-01
影响因子：
2.9
通讯作者：
CHAPA, FJ
中科院分区：
文献类型：
--
作者：
REAL, MD;FERRE, J;CHAPA, FJ
为了研究果蝇提取物葡萄糖基化外源底物的能力，我们已经开发了一种快速和灵敏的方法，用于检测UDP-葡萄糖基转移酶活性使用4-硝基苯酚，1-萘酚，或2-萘酚作为底物。使用高效液相色谱法分离和定量反应产物，允许检测产生低至1 pmol 2-萘酚葡糖苷（荧光检测）或16 pmol 4-硝基苯酚葡糖苷（吸光度检测）的活性。果蝇酶对1-萘酚或2-萘酚的亲和力（Km = 4 μm）比对4-硝基苯酚和UDP-葡萄糖的亲和力（Km = 1 mm）高250倍。
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°C and alkaline pH. The affinity of the Drosophila enzyme was 250-fold higher for 1-naphthol or 2-naphthol (Km≈ 4 μm) than for 4-nitrophenol and UDP-glucose (Km≈ 1 mm).