title: 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 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.
authors.affiliation: CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.
authors.forename: André C
authors.initials: AC
authors.lastname: Müller
authors.affiliation: CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.
authors.forename: Roberto
authors.initials: R
authors.lastname: Giambruno
authors.affiliation: CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.
authors.forename: Juliane
authors.initials: J
authors.lastname: Weißer
authors.affiliation: CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.
authors.forename: Peter
authors.initials: P
authors.lastname: Májek
authors.affiliation: University of Zurich, Department of Chemistry, Zurich, Switzerland.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Hofer
authors.affiliation: CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.
authors.forename: Johannes W
authors.initials: JW
authors.lastname: Bigenzahn
authors.affiliation: CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.
authors.forename: Giulio
authors.initials: G
authors.lastname: Superti-Furga
authors.affiliation: University of Zurich, Department of Chemistry, Zurich, Switzerland.
authors.forename: Henning J
authors.initials: HJ
authors.lastname: Jessen
authors.affiliation: CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria.
authors.forename: Keiryn L
authors.initials: KL
authors.lastname: Bennett
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000032:analysis* / Q000737:chemistry*; D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D053487:DEAD-box RNA Helicases / Q000737:chemistry; D057809:HEK293 Cells; D006801:Humans; D007700:Kinetics; D013058:Mass Spectrometry; D010103:Oxygen Isotopes / Q000737:chemistry; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: C513733:ABI1 protein, human; D048868:Adaptor Proteins, Signal Transducing; D003598:Cytoskeletal Proteins; D010103:Oxygen Isotopes; C000615259:Oxygen-18; D010455:Peptides; D000255:Adenosine Triphosphate; D011494:Protein Kinases; C119332:DDX3X protein, human; D053487:DEAD-box RNA Helicases