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Functional display of heterotetrameric human protein kinase CK2 on Escherichia coli: a novel tool for drug discovery

Background: Human protein kinase CK2 represents a novel therapeutic target for neoplastic diseases. Inhibitors are in need to explore the druggability and the therapeutic options of this enzyme. A bottleneck in the search for new inhibitors is the availability of the target for testing. Therefore an assay was developed to provide easy access to CK2 for discovery of novel inhibitors. Results: Autodisplay was used to present human CK2 on the surface of Escherichia coli. Heterotetrameric CK2 consists of two subunits, α and β, which were displayed individually on the surface. Co-display of CK2α and CK2β on the cell surface led to the formation of functional holoenzyme, as demonstrated by NaCl dependency of enzymatic activity, which differs from that of the catalytic subunit CK2α without β. In addition interaction of CK2α and CK2β at the cell surface was confirmed by co-immunoprecipitation assays. Surface displayed CK2 holoenzyme enabled an easy IC50 value determination. The IC50 values for the known CK2 inhibitors TBB and Silmitasertib were determined to be 50 and 3.3 nM, respectively. Conclusion: Surface-displayed CK2α and CK2β assembled on the cell surface of E. coli to an active tetrameric holoenzyme. The whole-cell CK2 autodisplay assay as developed is suitable for inhibition studies. Furthermore, it can be used to determine quantitative CK2 inhibition data such as IC50 values. In summary, this is the first report on the functional surface display of a heterotetrameric enzyme on E. coli.

Titel: Functional display of heterotetrameric human protein kinase CK2 on Escherichia coli: a novel tool for drug discovery
Verfasser: Gratz, Andreas GND
Bollacke, Andre GND
Stephan, Sara GND
Nienberg, Christian
Borgne, Marc Le
Götz , Claudia
Jose, Joachim
Organisation: FB 12: Chemie und Pharmazie
Dokumenttyp: Artikel
Medientyp: Text
Erscheinungsdatum: 03.06.2015
Publikation in MIAMI: 23.06.2015
Datum der letzten Änderung: 23.05.2019
Quelle: Microbial Cell Factories 14 (2015) 74, 1-13
Schlagwörter: Assay; Autodisplay; CK2; Heterotetramer; Protein kinase; Screening; Surface display; Drug discovery
Fachgebiete: Chemie
Lizenz: CC BY 4.0
Sprache: Englisch
Anmerkungen: Finanziert durch den Open-Access-Publikationsfonds 2015/2016 der Westfälischen Wilhelms-Universität Münster (WWU Münster).
Format: PDF-Dokument
URN: urn:nbn:de:hbz:6-49229555362
DOI: 10.1186/s12934-015-0263-z
ISSN: 1475-2859