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Identifying Kinase Substrates via a Heavy ATP Kinase Assay and Quantitative Mass Spectrometry.

  • Müller AC CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.
  • Giambruno R CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.
  • Weißer J CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.
  • Májek P CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.
  • Hofer A University of Zurich, Department of Chemistry, Zurich, Switzerland.
  • Bigenzahn JW CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.
  • Superti-Furga G CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.
  • Jessen HJ University of Zurich, Department of Chemistry, Zurich, Switzerland.
  • Bennett KL CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.
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  • 2016-06-28
Published in:
  • Scientific reports. - 2016
Adaptor Proteins, Signal Transducing
Adenosine Triphosphate
Amino Acid Sequence
Chromatography, High Pressure Liquid
Cytoskeletal Proteins
DEAD-box RNA Helicases
HEK293 Cells
Humans
Kinetics
Mass Spectrometry
Oxygen Isotopes
Peptides
Phosphorylation
Protein Kinases
Substrate Specificity
English Mass spectrometry-based in vitro kinase screens play an essential role in the discovery of kinase substrates, however, many suffer from biological and technical noise or necessitate genetically-altered enzyme-cofactor systems. We describe a method that combines stable γ-[(18)O2]-ATP with classical in vitro kinase assays within a contemporary quantitative proteomic workflow. Our approach improved detection of known substrates of the non-receptor tyrosine kinase ABL1; and identified potential, new in vitro substrates.
Language
  • English
Open access status
gold
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Persistent URL
https://sonar.ch/global/documents/96901
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