Issue 6, 2016

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.

Graphical abstract: Photo-crosslinking of clinically relevant kinases using H89-derived photo-affinity probes

Supplementary files

Article information

Article type
Paper
Submitted
06 Сәу. 2016
Accepted
19 Сәу. 2016
First published
19 Сәу. 2016

Mol. BioSyst., 2016,12, 1809-1817

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

S. C. Stolze, N. Liu, R. H. Wijdeven, A. W. Tuin, A. M. C. H. van den Nieuwendijk, B. I. Florea, M. van der Stelt, G. A. van der Marel, J. J. Neefjes and H. S. Overkleeft, Mol. BioSyst., 2016, 12, 1809 DOI: 10.1039/C6MB00257A

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