A Multiplexed Nmr-Reporter Approach To Measure Cellular Kinase And Phosphatase Activities In Real-Time
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY(2015)
摘要
Cell signaling is governed by dynamic changes in kinase and phosphatase activities, which are difficult to assess with discontinuous readout methods. Here, we introduce an NMR-based reporter approach to directly identify active kinases and phosphatases in complex physiological environments such as cell lysates and to measure their individual activities in a semicontinuous fashion. Multiplexed NMR profiling of reporter phosphorylation states provides unique advantages for kinase inhibitor studies and reveals reversible modulations of cellular enzyme activities under different metabolic conditions.
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关键词
phosphatase activities,cellular kinase,nmr-reporter,real-time
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