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Thrombopoietin receptor-based protein-protein interaction screening (THROPPIS)
Makiko Horikawa1, Yosuke Kakiuchi1, Daiki Kashima2
1Department of Bioengineering, Graduate School of Engineering, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.
Biotechnology and Bioengineering
|October 28, 2021
Summary
Researchers developed a new system (THROPPIS) to detect protein-protein interactions (PPIs) using the thrombopoietin receptor. This system enhances PPI detection sensitivity and can be extended to other receptors for drug discovery.
Area of Science:
- Molecular Biology
- Cellular Biology
- Biochemistry
Background:
- Protein-protein interactions (PPIs) are crucial for cellular functions and are targets for drug discovery.
- Existing PPI detection systems, like the c-kit-based PPI screening (KIPPIS), have limitations.
- The potential of using other receptors for PPI detection remains largely unexplored.
Purpose of the Study:
- To develop a novel mammalian cytosolic protein-protein interaction detection system using the thrombopoietin receptor.
- To enhance the sensitivity and versatility of this new system, named thrombopoietin receptor-based PPI screening (THROPPIS).
- To demonstrate the applicability of the THROPPIS system with different receptor tyrosine kinases.
Main Methods:
- Engineered chimeric receptors by fusing proteins of interest with the thrombopoietin receptor.
- Investigated strategies to improve system sensitivity, including receptor localization and addition of helper modules.
- Validated the system by detecting known peptide-domain interactions.
Main Results:
- The thrombopoietin receptor-based PPI screening (THROPPIS) system was successfully developed.
- Non-localized chimeric receptors exhibited optimal performance.
- The addition of a helper module significantly enhanced detection sensitivity.
- Successfully detected 5 peptide-domain interactions, showcasing THROPPIS versatility.
- Demonstrated PPI detection using insulin receptor and epidermal growth factor receptor signaling domains.
Conclusions:
- The developed THROPPIS system offers a sensitive and versatile method for detecting protein-protein interactions in mammalian cells.
- The system's modularity allows for adaptation to various receptors, expanding its utility in biological research and drug discovery.
- THROPPIS represents a valuable advancement in the field of PPI detection technologies.
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