- Identifying Kinase Substrates via a Heavy ATP Kinase Assay and Quantitative Mass Spectrometry.
Identifying Kinase Substrates via a Heavy ATP Kinase Assay and Quantitative Mass Spectrometry.
Scientific reports (2016-06-28)
André C Müller, Roberto Giambruno, Juliane Weißer, Peter Májek, Alexandre Hofer, Johannes W Bigenzahn, Giulio Superti-Furga, Henning J Jessen, Keiryn L Bennett
PMID27346722
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.