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Sci Rep. 2016 Jun 27;6:28107. doi: 10.1038/srep28107.

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

Author information

1
CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.
2
University of Zurich, Department of Chemistry, Zurich, Switzerland.

Abstract

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.

PMID:
27346722
PMCID:
PMC4921819
DOI:
10.1038/srep28107
[Indexed for MEDLINE]
Free PMC Article

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