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    • 7. 发明授权
    • Method for quantitative analysis for prolyl hydroxylase activity using fluorescence polarization
    • 使用荧光偏振定量分析脯氨酰羟化酶活性的方法
    • US07691966B2
    • 2010-04-06
    • US11164813
    • 2005-12-06
    • Eun Gyeong YangHyun Ju ChoHyunsung Park
    • Eun Gyeong YangHyun Ju ChoHyunsung Park
    • C07K14/00C12Q1/00C12Q1/30
    • G01N33/6875G01N33/6872G01N2500/00
    • The present invention relates to a method for analyzing the interaction between HIF-1 peptide and VBC protein using fluorescence polarization, more precisely, a method for quantitative analysis of formation of HIF-1-VBC protein complex which is composed of the steps of 1) preparing a fluorescent probe by attaching a fluorescein to hydroxyproline containing HIF-1 peptide; 2) reacting the fluorescent probe with VBC protein; and 3) measuring the fluorescence polarization of the above reactant and then comparing the fluorescence polarization with that of the fluorescent probe itself to investigate the changes of fluorescence polarization; a method for screening an inhibitor of the binding of HIF-1 peptide and VBC protein using the above method; and a method for analyzing the activity of prolyl hydroxylase using the above method. The method of the present invention enables simple analysis of the interaction between HIF-1 peptide and VBC protein by observing changes of fluorescence polarization and thus, it can be effectively used for high-speed screening using a well plate.
    • 本发明涉及使用荧光偏振分析HIF-1肽与VBC蛋白之间相互作用的方法,更确切地说,涉及HIF-1-VBC蛋白复合物形成定量分析方法,该方法由以下步骤组成:1) 通过将荧光素附着到含有HIF-1肽的羟脯氨酸上制备荧光探针; 2)使荧光探针与VBC蛋白反应; 和3)测量上述反应物的荧光偏振,然后将荧光偏振与荧光探针本身的荧光偏振进行比较,以研究荧光偏振的变化; 使用上述方法筛选HIF-1肽和VBC蛋白的结合抑制剂的方法; 以及使用上述方法分析脯氨酰羟化酶的活性的方法。 通过观察荧光偏振的变化,本发明的方法能够简单地分析HIF-1肽与VBC蛋白之间的相互作用,因此可以有效地用于使用孔板的高速筛选。