Gratz, Andreas; Bollacke, Andre; Stephan, Sara; Nienberg, Christian; Borgne, Marc Le; Götz , Claudia; Jose, Joachim: Functional display of heterotetrameric human protein kinase CK2 on Escherichia coli: a novel tool for drug discovery
Inhalt
- Bacterial strains and culture conditions
- Design of autotransporter fusion genes and expression plasmids
- Outer membrane protein preparation and trypsin accessibility
- SDS-PAGE and Western blot
- Co-immunoprecipitation
- Radiometric whole-cell CK2 activity
- Whole-cell CK2 activity assay and inhibition testing by capillary electrophoresis
- Influence of NaCl on the enzymatic activity of surface displayed CK2α and CK2 holoenzyme
- Results
- Autodisplay of CK2 subunits α and β
- Interaction of CK2α and CK2β on the cell surface
- Kinase activity of E. coli cells displaying CK2α and CK2α + CK2β
- CK2 holoenzyme formation on the cell surface
- Inhibitor testing
- Discussion
- Conclusion
- Authors’ contributions
- Received: 20 March 2015 Accepted: 11 May 2015References