Next Article in Journal
Strategies to Overcome Heparins’ Low Oral Bioavailability
Next Article in Special Issue
Convergent Synthesis of Two Fluorescent Ebselen-Coumarin Heterodimers
Previous Article in Journal
Announcing the 2016 Pharmaceuticals Travel Award for Young Investigators
Previous Article in Special Issue
Synthesis and Pharmacological Properties of Novel Esters Based on Monocyclic Terpenes and GABA
Article Menu

Export Article

Open AccessFeature PaperArticle
Pharmaceuticals 2016, 9(3), 36; doi:10.3390/ph9030036

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

1
Institut für Pharmazeutische und Medizinische Chemie, PharmaCampus, Westfälische Wilhelms-Universität Münster, Corrensstraße 48, D-48149 Münster, Germany
2
Institut für Biochemie, Westfälische Wilhelms-Universität Münster, Wilhelm-Klemm-Straße 2, D-48149 Münster, Germany
The best presentation at the 1st International Electronic Conference on Medicinal Chemistry.
*
Author to whom correspondence should be addressed.
Academic Editor: Jean Jacques Vanden Eynde
Received: 19 May 2016 / Revised: 15 June 2016 / Accepted: 17 June 2016 / Published: 27 June 2016
View Full-Text   |   Download PDF [4407 KB, uploaded 27 June 2016]   |  

Abstract

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
Figures

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

Scifeed alert for new publications

Never miss any articles matching your research from any publisher
  • Get alerts for new papers matching your research
  • Find out the new papers from selected authors
  • Updated daily for 49'000+ journals and 6000+ publishers
  • Define your Scifeed now

SciFeed Share & Cite This Article

MDPI and ACS Style

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.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Pharmaceuticals EISSN 1424-8247 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top