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    • 6. 发明申请
    • METHODS FOR IDENTIFYING INHIBITORS OF STEROL 14-alpha-DEMETHYLASE
    • 用于鉴定甾体14-α-脱乙酰酶抑制剂的方法
    • WO2005033664A2
    • 2005-04-14
    • PCT/US2004032633
    • 2004-10-01
    • ICORIA INCASLAMKHAN ABUBAKRGUO LININGRICE JOHN W
    • ASLAMKHAN ABUBAKRGUO LININGRICE JOHN W
    • C12Q1/26C12Q1/32G01N
    • C12Q1/26C12Q1/32G01N2500/00
    • In the methods of the present invention, the change in absorbance of an obtusifoliol 14alpha-demethylase (OBT-DM) enzyme upon binding of an inhibitor is used to advantage for the identification of inhibitors of OBT-DM activity. Rather than measuring the absorbance of an OBT-DM/inhibitor complex over a spectrum of wavelengths as described previously, the present invention discloses methods for identifying type II inhibitors of OBT-DM by monitoring the absorbance only at 413nm and 432 nm. The methods of the invention enable the concurrent testing of multiple compounds using a high throughput format such as with 96- or 384-well plates. The OBT-DM polypeptides of the invention include plant, fungal and human OBT-DM polypeptides, and in particular, Arabidopsis thaliana OBT-DM polypeptide.
    • 在本发明的方法中,使用抑制剂结合时,obtusifoliol14α-去甲基化酶(OBT-DM)的吸光度变化有利于鉴定OBT-DM活性的抑制剂。 本发明不是如上所述在波长谱上测量OBT-DM /抑制剂复合物的吸光度,而是通过仅在413nm和432nm监测吸光度来公开鉴定OBT-DM的II型抑制剂的方法。 本发明的方法使得能够使用高通量形式(例如96孔或384孔板)同时测试多种化合物。 本发明的OBT-DM多肽包括植物,真菌和人类OBT-DM多肽,尤其是拟南芥OBT-DM多肽。