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Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach
11:11

Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach

Published on: February 21, 2019

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Unexpected CK2β-antagonistic functionality of bisubstrate inhibitors targeting protein kinase CK2

Markus Pietsch1, Kaido Viht2, Alexander Schnitzler3

  • 1Institut II für Pharmakologie, Zentrum für Pharmakologie, Medizinische Fakultät, Universität zu Köln, Gleueler Str. 24, D-50931 Köln, Germany.

Bioorganic Chemistry
|February 15, 2020
PubMed

Insights

A novel inhibitor, ARC-3140, targets the protein kinase CK2 holoenzyme by binding to both ATP and substrate sites. This bisubstrate inhibitor significantly impacts CK2α/CK2β interactions, offering a new avenue for cancer drug development.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Drug Discovery

Background:

  • Protein kinase CK2 (CK2) is a heterotetrameric enzyme implicated in cancer.
  • CK2's structure involves catalytic (CK2α) and regulatory (CK2β) subunits.
  • Targeting CK2 is a promising strategy for cancer therapy.

Purpose of the Study:

  • To introduce ARC-3140, a novel bisubstrate inhibitor of CK2.
  • To elucidate the mechanism of ARC-3140's interaction with CK2.
  • To evaluate ARC-3140 as a lead compound for cancer drug development.

Main Methods:

  • X-ray crystallography to determine the structure of CK2 in complex with ARC-3140.
  • Microscale thermophoresis and fluorescence anisotropy to study protein-protein interactions.
  • Biochemical assays to assess inhibitor affinity and mechanism.

Main Results:

  • ARC-3140 exhibits extremely high affinity (Ki = 84 pM) for CK2.
  • Crystal structure reveals ARC-3140 binding at the ATP and substrate sites, and influencing CK2α/CK2β interfaces.
  • ARC-3140 and ARC-1502 disrupt the bipartite interaction between CK2α and CK2β subunits.

Conclusions:

  • ARC-3140 is a potent bisubstrate inhibitor of CK2.
  • ARC-3140 uniquely interferes with both sub-interfaces of the CK2α/CK2β interaction.
  • ARC-3140 represents a promising lead for developing novel cancer therapeutics targeting CK2 quaternary structure.
Keywords:
Bisubstrate inhibitorCK2β antagonistsCapillary electrophoresis-based kinase assayFluorescence anisotropy assayMicroscale thermophoresisProtein kinase CK2Protein/protein interactionX-ray crystallography

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