Engineering mono- and multi-valent inhibitors on a modular scaffold
| Autor: | Aurora Diamante, Piyush K. Chaturbedy, Pamela J. E. Rowling, Janet R. Kumita, Rohan S. Eapen, Stephen H. McLaughlin, Marc de la Roche, Albert Perez-Riba, Laura S. Itzhaki |
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| Přispěvatelé: | Kumita, Janet R [0000-0002-3887-4964], Eapen, Rohan S [0000-0001-7269-3633], Perez-Riba, Albert [0000-0002-4659-0320], Itzhaki, Laura S [0000-0001-6504-2576], Apollo - University of Cambridge Repository |
| Rok vydání: | 2021 |
| Předmět: |
FOS: Nanotechnology
0303 health sciences Scaffold Cellular activity 34 Chemical Sciences Chemistry business.industry Wnt signaling pathway Bioengineering General Chemistry Modular architecture Modular design 010402 general chemistry 01 natural sciences Small molecule 0104 chemical sciences 03 medical and health sciences Tetratricopeptide Biophysics Nanotechnology Generic health relevance business 030304 developmental biology Macromolecule |
| Popis: | Here we exploit the simple, ultra-stable, modular architecture of consensus-designed tetratricopeptide repeat proteins (CTPRs) to create a platform capable of displaying both single as well as multiple functions and with diverse programmable geometrical arrangements by grafting non-helical short linear binding motifs (SLiMs) onto the loops between adjacent repeats. As proof of concept, we built synthetic CTPRs to bind and inhibit the human tankyrase proteins (hTNKS), which play a key role in Wnt signaling and are upregulated in cancer. A series of mono-valent and multi-valent hTNKS binders was assembled. To fully exploit the modular scaffold and to further diversify the multi-valent geometry, we engineered the binding modules with two different formats, one monomeric and the other trimeric. We show that the designed proteins are stable, correctly folded and capable of binding to and inhibiting the cellular activity of hTNKS leading to downregulation of the Wnt pathway. Multivalency in both the CTPR protein arrays and the hTNKS target results in the formation of large macromolecular assemblies, which can be visualized both in vitro and in the cell. When delivered into the cell by nanoparticle encapsulation, the multivalent CTPR proteins displayed exceptional activity. They are able to inhibit Wnt signaling where small molecule inhibitors have failed to date. Our results point to the tremendous potential of the CTPR platform to exploit a range of SLiMs and assemble synthetic binding molecules with built-in multivalent capabilities and precise, pre-programmed geometries. |
| Databáze: | OpenAIRE |
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