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Updated: Jan 1, 2026
Identification of Kinase-substrate Pairs Using High Throughput Screening
Published on: August 29, 2015
Fragment-based screening identifies molecules targeting the substrate-binding ankyrin repeat domains of tankyrase
Katie Pollock1,2,3, Manjuan Liu2, Mariola Zaleska1
1Divisions of Structural Biology & Cancer Biology, The Institute of Cancer Research (ICR), London, SW7 3RP, United Kingdom.
Abstract:
The PARP enzyme and scaffolding protein tankyrase (TNKS, TNKS2) uses its ankyrin repeat clusters (ARCs) to bind a wide range of proteins and thereby controls diverse cellular functions. A number of these are implicated in cancer-relevant processes, including Wnt/β-catenin signalling, Hippo signalling and telomere maintenance. The ARCs recognise a conserved tankyrase-binding peptide motif (TBM). All currently available tankyrase inhibitors target the catalytic domain and inhibit tankyrase's poly(ADP-ribosyl)ation function. However, there is emerging evidence that catalysis-independent "scaffolding" mechanisms contribute to tankyrase function. Here we report a fragment-based screening programme against tankyrase ARC domains, using a combination of biophysical assays, including differential scanning fluorimetry (DSF) and nuclear magnetic resonance (NMR) spectroscopy. We identify fragment molecules that will serve as starting points for the development of tankyrase substrate binding antagonists. Such compounds will enable probing the scaffolding functions of tankyrase, and may, in the future, provide potential alternative therapeutic approaches to inhibiting tankyrase activity in cancer and other conditions.
Insights
Researchers identified new ways to target the tankyrase (TNKS) protein, which is involved in cancer. This study focused on blocking its scaffolding function, offering potential new cancer therapies.
Area of Science:
- Biochemistry
- Molecular Biology
- Drug Discovery
Background:
- Tankyrase (TNKS, TNKS2) is a PARP enzyme and scaffolding protein crucial for cellular functions.
- Its ankyrin repeat clusters (ARCs) bind proteins, regulating cancer-relevant pathways like Wnt/β-catenin signaling, Hippo signaling, and telomere maintenance.
- Existing tankyrase inhibitors target the catalytic domain, but scaffolding functions are increasingly recognized.
Purpose of the Study:
- To identify novel inhibitors targeting the scaffolding function of tankyrase ARCs.
- To develop tools for probing tankyrase's catalysis-independent roles.
- To explore alternative therapeutic strategies for tankyrase inhibition in cancer.
Main Methods:
- Fragment-based screening against tankyrase ARC domains.
- Utilized biophysical assays, including differential scanning fluorimetry (DSF).
- Employed nuclear magnetic resonance (NMR) spectroscopy.
Main Results:
- Identified fragment molecules that bind to tankyrase ARC domains.
- These fragments serve as starting points for developing tankyrase substrate binding antagonists.
- Demonstrated a novel approach to target tankyrase's scaffolding activity.
Conclusions:
- The identified fragments can be developed into inhibitors of tankyrase scaffolding function.
- This approach offers a new avenue for understanding and targeting tankyrase in cancer.
- Potential for developing alternative therapeutic strategies beyond catalytic inhibition.
