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

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 ?-[(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.

SUBMITTER: Muller AC

PROVIDER: S-EPMC4921819 | biostudies-literature | 2016 Jun

REPOSITORIES: biostudies-literature

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

Müller André C AC Giambruno Roberto R Weißer Juliane J Májek Peter P Hofer Alexandre A Bigenzahn Johannes W JW Superti-Furga Giulio G Jessen Henning J HJ Bennett Keiryn L KL

Scientific reports 20160627

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 ...[more]

PMID: 27346722

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