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

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)O(2)]-ATP with classical...

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Detalles Bibliográficos
Autores principales: Müller, André C., Giambruno, Roberto, Weißer, Juliane, Májek, Peter, Hofer, Alexandre, Bigenzahn, Johannes W., Superti-Furga, Giulio, Jessen, Henning J., Bennett, Keiryn L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4921819/
https://www.ncbi.nlm.nih.gov/pubmed/27346722
http://dx.doi.org/10.1038/srep28107
Descripción
Sumario: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)O(2)]-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.

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