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Open AccessFeature PaperArticle

Site-Specific Labeling of Protein Kinase CK2: Combining Surface Display and Click Chemistry for Drug Discovery Applications

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Institut für Pharmazeutische und Medizinische Chemie, PharmaCampus, Westfälische Wilhelms-Universität Münster, Corrensstraße 48, D-48149 Münster, Germany
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Institut für Biochemie, Westfälische Wilhelms-Universität Münster, Wilhelm-Klemm-Straße 2, D-48149 Münster, Germany
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Author to whom correspondence should be addressed.
The best presentation at the 1st International Electronic Conference on Medicinal Chemistry.
Academic Editor: Jean Jacques Vanden Eynde
Pharmaceuticals 2016, 9(3), 36; https://doi.org/10.3390/ph9030036
Received: 19 May 2016 / Revised: 15 June 2016 / Accepted: 17 June 2016 / Published: 27 June 2016
Human CK2 is a heterotetrameric constitutively active serine/threonine protein kinase and is an emerging target in current anti-cancer drug discovery. The kinase is composed of two catalytic CK2α subunits and two regulatory CK2β subunits. In order to establish an assay to identify protein-protein-interaction inhibitors (PPI) of the CK2α/CK2β interface, a bioorthogonal click reaction was used to modify the protein kinase α-subunit with a fluorophore. By expanding the genetic code, the unnatural amino acid para azidophenylalanine (pAzF) could be incorporated into CK2α. Performing the SPAAC click reaction (Strain-Promoted Azide-Alkyne Cycloaddition) by the use of a dibenzylcyclooctyne-fluorophore (DBCO-fluorophore) led to a specifically labeled human protein kinase CK2α. This site-specific labeling does not impair the phosphorylation activity of CK2, which was evaluated by capillary electrophoresis. Furthermore a dissociation constant (KD) of 631 ± 86.2 nM was determined for the substrate αS1-casein towards CK2α. This labeling strategy was also applied to CK2β subunit on Escherichia coli, indicating the site-specific modifications of proteins on the bacterial cell surface when displayed by Autodisplay. View Full-Text
Keywords: CK2; kinase; Autodisplay; click chemistry; unnatural amino acid; bioorthogonal; labeling; drug discovery; protein-protein interaction CK2; kinase; Autodisplay; click chemistry; unnatural amino acid; bioorthogonal; labeling; drug discovery; protein-protein interaction
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Nienberg, C.; Retterath, A.; Becher, K.-S.; Saenger, T.; Mootz, H.D.; Jose, J. Site-Specific Labeling of Protein Kinase CK2: Combining Surface Display and Click Chemistry for Drug Discovery Applications. Pharmaceuticals 2016, 9, 36.

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