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1. 发明申请

WO0125477A3 KINASE AND PHOSPHATASE ASSAYS 审中-公开
标题翻译：激酶和磷酸酶测定
公开(公告)号：WO0125477A3
公开(公告)日：2002-01-31
申请号：PCT/GB0003736
申请日：2000-09-29
申请人： BIOFOCUS DISCOVERY LTD , FREARSON JULIE ANN
发明人： FREARSON JULIE ANN
IPC分类号： C12Q1/42 , C12Q1/48 , G01N33/573
CPC分类号： C12Q1/485 , C12Q1/42 , G01N2500/00
摘要： Methods for screening for modulators of kinase or phosphatase activity are described. The methods comprise: a) providing a serine or threonine kinase and a substrate which can be phosphorylated by the kinase; b) contacting the kinase with its substrate in the presence and absence of a candidate modulator of the activity of the kinase under conditions which permit phosphorylation of the substrate by the kinase; c) contacting the substrates treated according to step (b) with a reporter, excluding a natural antibody, which binds phosphorylated substrate with higher affinity than unphosphorylated substrate; and d) comparing binding of the reporter to the substrate treated according to step (b) in the presence of the candidate modulator with binding of the reporter to the substrate treated according to step (b) in the absence of the candidate modulator. Examples of reporters for use in such assays include 14-3-3 proteins and WW domain containing proteins.
摘要翻译：描述了筛选激酶或磷酸酶活性调节剂的方法。所述方法包括：a）提供可由激酶磷酸化的丝氨酸或苏氨酸激酶和底物; b）在允许由激酶磷酸化底物的条件下，在存在和不存在激酶活性的候选调节剂的情况下，使激酶与其底物接触; c）使根据步骤（b）处理的底物与不含天然抗体的报告基团接触，所述报告物以比非磷酸化底物更高的亲和力结合磷酸化底物; 以及d）比较在候选调节剂存在下报告基团与根据步骤（b）处理的底物的结合，所述候选调节剂在不存在候选调节剂的情况下结合报道基因与根据步骤（b）处理的底物结合。用于此类测定的记者的实例包括14-3-3蛋白质和含有WW结构域的蛋白质。

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