title: Photo-crosslinking of clinically relevant kinases using H89-derived photo-affinity probes.
abstract: The profiling of kinases using established proteomics techniques is hampered by their non-covalent mode-of-action. One way to overcome this caveat is the use of probes featuring photo-labelling groups that can be activated by UV irradiation to generate a reactive species that will establish a covalent bond to the enzyme. In this study we have used the well-known kinase inhibitor H89 as a lead for the development of probes for the affinity-based profiling of clinically relevant kinases. A labelling protocol was established for recombinant kinases and more complex protein mixtures using gel-based techniques. We also show that the probes act in a competitive manner with other kinase inhibitors.
authors.affiliation: Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands. h.s.overkleeft@chem.leidenuniv.nl.
authors.forename: Sara C
authors.initials: SC
authors.lastname: Stolze
authors.affiliation: Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands. h.s.overkleeft@chem.leidenuniv.nl.
authors.forename: Nora
authors.initials: N
authors.lastname: Liu
authors.affiliation: Division of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.
authors.forename: Ruud H
authors.initials: RH
authors.lastname: Wijdeven
authors.affiliation: Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands. h.s.overkleeft@chem.leidenuniv.nl.
authors.forename: Adriaan W
authors.initials: AW
authors.lastname: Tuin
authors.affiliation: Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands. h.s.overkleeft@chem.leidenuniv.nl.
authors.forename: Adrianus M C H
authors.initials: AM
authors.lastname: van den Nieuwendijk
authors.affiliation: Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands. h.s.overkleeft@chem.leidenuniv.nl.
authors.forename: Bogdan I
authors.initials: BI
authors.lastname: Florea
authors.affiliation: Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands. h.s.overkleeft@chem.leidenuniv.nl.
authors.forename: Mario
authors.initials: M
authors.lastname: van der Stelt
authors.affiliation: Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands. h.s.overkleeft@chem.leidenuniv.nl.
authors.forename: Gijsbert A
authors.initials: GA
authors.lastname: van der Marel
authors.affiliation: Division of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.
authors.forename: Jacques J
authors.initials: JJ
authors.lastname: Neefjes
authors.affiliation: Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands. h.s.overkleeft@chem.leidenuniv.nl.
authors.forename: Herman S
authors.initials: HS
authors.lastname: Overkleeft
mesh_terms: D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry / Q000378:metabolism; D004789:Enzyme Activation; D031541:Fluorescence Resonance Energy Transfer; D007546:Isoquinolines / Q000737:chemistry*; D007700:Kinetics; D015335:Molecular Probes / Q000737:chemistry*; D015394:Molecular Structure; D010770:Phosphotransferases / Q000737:chemistry* / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000737:chemistry / Q000378:metabolism; D013194:Staining and Labeling; D013449:Sulfonamides / Q000737:chemistry*; D014466:Ultraviolet Rays
chemical_list: D007546:Isoquinolines; D015335:Molecular Probes; D013449:Sulfonamides; D010770:Phosphotransferases; D051057:Proto-Oncogene Proteins c-akt; D017868:Cyclic AMP-Dependent Protein Kinases; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide