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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Metal ion-mediated polymer superquenching for highly sensitive detection of kinase and phosphatase activities
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Metal ion-mediated polymer superquenching for highly sensitive detection of kinase and phosphatase activities

机译：金属离子介导的聚合物超淬灭，可高度灵敏地检测激酶和磷酸酶活性

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摘要

An assay technology for high-throughput screening of kinase and phosphatase activities is introduced. The format is based upon superquenching of fluorescent-conjugated polymers by dye-labeled kinase/phosphatase peptide substrates. The sensor platform is composed of highly fluorescent-conjugated polyelectrolytes colocated with the phosphate coordinating metal ion gallium on microspheres. Phosphorylated peptide substrates containing a quencher bind specifically to the metal ions by means of phosphate groups, resulting in quench of polymer fluorescence. The modulation of fluorescence signal is proportional to kinase or phosphatase activity and is monitored as a turn-off or turn-on signal, respectively. The assay is homogeneous and simple and can be run either as an endpoint measurement or in a kinetic mode. The assay meets the sensitivity required for high-throughput screening of kinase or phosphatase inhibitors and is a valuable tool for drug discovery. A modified version of the assay allows for the detection of protein phosphorylation.
机译：介绍了用于高通量筛选激酶和磷酸酶活性的测定技术。该形式基于染料标记的激酶/磷酸酶肽底物对荧光共轭聚合物的超猝灭。传感器平台由与微球上的磷酸盐配位金属离子镓共存的高荧光共轭聚电解质组成。含有猝灭剂的磷酸化肽底物通过磷酸基团与金属离子特异性结合，从而导致聚合物荧光猝灭。荧光信号的调节与激酶或磷酸酶活性成正比，分别作为关闭或打开信号进行监控。该测定是均一且简单的，并且可以作为终点测量或以动力学模式进行。该测定满足激酶或磷酸酶抑制剂高通量筛选所需的灵敏度，并且是发现药物的宝贵工具。该测定的修改版本允许检测蛋白质磷酸化。

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