磷酸酶
丝氨酸
蛋白质酪氨酸磷酸酶
苏氨酸
生物化学
酪氨酸
化学
DUSP6型
酶
磷酸化
分子生物学
生物
蛋白磷酸酶2
作者
Christina Pastula,Iain Johnson,Joseph M. Beechem,Wayne F. Patton
出处
期刊:Combinatorial Chemistry & High Throughput Screening
[Bentham Science]
日期:2003-06-01
卷期号:6 (4): 341-346
被引量:18
标识
DOI:10.2174/138620703106298590
摘要
A number of aromatic substrates were evaluated for their ability to detect tyrosine phosphatase and serine / threonine phosphatase activity. Results demonstrated that the fluorinated coumarin DiFMUP is the most sensitive substrate for detecting LAR and PP-2A activity. Using this substrate, selective high-throughput screening assays for serine / threonine and tyrosine phosphatases were developed. Specific inhibitor cocktails were added to each assay to limit the activity of other phosphatases. LAR, CD-45, and PTP-1B all rapidly hydrolyze DiFMUP in the tyrosine phosphatase assay. The activity of non-tyrosine phosphatases is less than 6% of the LAR activity. PP-1 and PP-2A are highly active in the serine / threonine phosphatase assay. Inhibition of LAR and PP-2A in these assays is demonstrated using known inhibitors. Both of these assays are sensitive, robust, kinetic assays that can be used to quantify enzyme activity. Keywords: fluorescence, microplate, tyrosine phosphatase, serine/threonine phosphatase, phosphatase inhibitor
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