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A multiplexed NMR-reporter approach to measure cellular kinase and phosphatase activities in real-time

Authors

  • R. Thongwichian
  • J. Kosten
  • U. Benary
  • H.M. Rose
  • M. Stuiver
  • F.X. Theillet
  • A. Dose
  • B. Koch
  • H. Yokoyama
  • D. Schwarzer
  • J. Wolf
  • P. Selenko

Journal

  • Journal of the American Chemical Society

Citation

  • J Am Chem Soc 137 (20): 6468-6471

Abstract

  • 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.


DOI

doi:10.1021/jacs.5b02987