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

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

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-in...

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Bibliographic Details
Published in:Pharmaceuticals (Basel, Switzerland) Vol. 9; no. 3; p. 36
Main Authors: Nienberg, Christian, Retterath, Anika, Becher, Kira-Sophie, Saenger, Thorsten, Mootz, Henning D, Jose, Joachim
Format: Journal Article
Language:English
Published: Switzerland MDPI AG 27-06-2016
MDPI
Subjects:
Amino acids
Autodisplay
bioorthogonal
CK2
click chemistry
drug discovery
E coli
kinase
Kinases
labeling
Pharmacology
protein-protein interaction
Proteins
R&D
Research & development
unnatural amino acid
kinase
drug discovery
labeling
Autodisplay
click chemistry
protein-protein interaction
unnatural amino acid
CK2
bioorthogonal
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Summary: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.
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ISSN:1424-8247
1424-8247
DOI:10.3390/ph9030036