Photo-crosslinking of clinically relevant kinases using H89-derived photo-affinity probes.

Autor: Stolze SC; Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands. h.s.overkleeft@chem.leidenuniv.nl., Liu N; Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands. h.s.overkleeft@chem.leidenuniv.nl., Wijdeven RH; Division of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands., Tuin AW; Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands. h.s.overkleeft@chem.leidenuniv.nl., van den Nieuwendijk AM; Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands. h.s.overkleeft@chem.leidenuniv.nl., Florea BI; Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands. h.s.overkleeft@chem.leidenuniv.nl., van der Stelt M; Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands. h.s.overkleeft@chem.leidenuniv.nl., van der Marel GA; Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands. h.s.overkleeft@chem.leidenuniv.nl., Neefjes JJ; Division of Cell Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands., Overkleeft HS; Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300 RA Leiden, The Netherlands. h.s.overkleeft@chem.leidenuniv.nl.
Jazyk: angličtina
Zdroj: Molecular bioSystems [Mol Biosyst] 2016 May 24; Vol. 12 (6), pp. 1809-17.
DOI: 10.1039/c6mb00257a
Abstrakt: 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.
Databáze: MEDLINE
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