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Updated: Oct 20, 2025
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In-vivo Detection of Protein-protein Interactions on Micro-patterned Surfaces
Published on: March 19, 2010
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EndoBind detects endogenous protein-protein interactions in real time
Anke Bill1, Sheryll Espinola2, Daniel Guthy3
1Novartis Institute for Biochemical Research, Oncology, Cambridge, MA, USA. anke.bill@novartis.com.
Communications Biology
|September 16, 2021
Summary
We developed two new methods, RT-Bind and EndoBind, to track protein interactions over time. These techniques allow for continuous, real-time monitoring of protein dimerization for over two days.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Protein-protein interactions are crucial for cellular functions.
- Monitoring these interactions in real-time, especially for endogenous proteins, remains challenging.
Purpose of the Study:
- To develop and validate high-throughput compatible methods for detecting protein-protein interactions.
- To enable temporal evaluation of dimer formation over extended durations.
- To allow continuous, real-time monitoring of endogenous protein-protein interactions.
Main Methods:
- Developed RT-Bind for ectopically expressed proteins and EndoBind for endogenously tagged proteins.
- Both methods are high-throughput compatible.
- Utilized Nrf2-KEAP1 and CRAF-KRAS-G12V interactions as examples.
Main Results:
- Demonstrated the ability to detect protein-protein interactions for over 2 days post-substrate addition.
- RT-Bind and EndoBind provide temporal evaluation of dimer formation.
- Enabled continuous monitoring of endogenous protein-protein interactions in real time.
Conclusions:
- RT-Bind and EndoBind are robust, high-throughput compatible methods for studying protein-protein interactions.
- These methods facilitate the real-time, continuous monitoring of dynamic protein dimerization.
- The developed techniques offer significant advantages for studying protein interaction kinetics.
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