Chemical proteomics and functional proteomics strategies for protein kinase inhibitor validation and protein kinase substrate identification: Applications to protein kinase CK2

Laszlo Gyenis , J. P. Turowec , Maria Bretner , D. W. Litchfield


Since protein kinases have been implicated in numerous human diseases, kinase inhibitors have emerged as promising therapeutic agents. Despite this promise, there has been a relative lag in the development of unbiased strategies to validate both inhibitor specificity and the ability to inhibit target activity within living cells. To overcome these limitations, our efforts have been focused on the development of systematic strategies that employ chemical and functional proteomics. We utilized these strategies to evaluate small molecule inhibitors of protein kinase CK2, a constitutively active kinase that has recently emerged as target for anti-cancer therapy in clinical trials. Our chemical proteomics strategies used ATP or CK2 inhibitors immobilized on sepharose beads together with mass spectrometry to capture and identify binding partners from cell extracts. These studies have verified that interactions between CK2 and its inhibitors occur in complex mixtures. However, in the case of CK2 inhibitors related to 4,5,6,7-tetrabromo-1H-benzotriazole (TBB), our work has also revealed off-targets for the inhibitors. To complement these studies, we devised functional proteomics approaches to identify proteins that exhibit decreases in phosphorylation when cells are treated with CK2 inhibitors. To identify and validate those proteins that are direct substrates for CK2, we have also employed mutants of CK2 with decreased inhibitor sensitivity. Overall, our studies have yielded systematic platforms for studying CK2 inhibitors which we believe will foster efforts to define the biological functions of CK2 and to rigorously investigate its potential as a candidate for molecular-targeted therapy. This article is part of a Special Issue entitled: Inhibitors of Protein Kinases (2012).
Author Laszlo Gyenis - [Western University]
Laszlo Gyenis,,
, J. P. Turowec - [Western University]
J. P. Turowec,,
, Maria Bretner (FC / IBC / DDTB)
Maria Bretner,,
- Department Of Drug Technology And Biotechnology
, D. W. Litchfield - [Western University]
D. W. Litchfield,,
Journal seriesBiochimica et Biophysica Acta (BBA) - Proteins and Proteomics, ISSN 1570-9639
Issue year2013
Publication size in sheets0.5
Keywords in EnglishCK2, Protein kinase inhibitor, TBB, Chemoproteomics, Functional proteomics, Inhibitor profiling
ASJC Classification1312 Molecular Biology; 1303 Biochemistry; 1304 Biophysics; 1602 Analytical Chemistry
Languageen angielski
Biochimica et Biophysica Acta.pdf 668.23 KB
Score (nominal)30
Score sourcejournalList
ScoreMinisterial score = 25.0, 01-02-2020, ArticleFromJournal
Ministerial score (2013-2016) = 30.0, 01-02-2020, ArticleFromJournal
Publication indicators Scopus Citations = 8; WoS Citations = 8; Scopus SNIP (Source Normalised Impact per Paper): 2013 = 1.148; WoS Impact Factor: 2013 = 3.191 (2) - 2013=3.282 (5)
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