Analyzing Protein Interactions in Living Cells Using Split-TEV Assays
Author Information
Author(s): Wehr Michael C, Reinecke Lisa, Botvinnik Anna, Rossner Moritz J
Primary Institution: Max-Planck-Institute of Experimental Medicine, Göttingen, Germany
Hypothesis
The split-TEV system can effectively quantify phosphorylation-dependent protein-protein interactions in living mammalian cells.
Conclusion
The split-TEV assays are effective for measuring phosphorylation-dependent and transient protein-protein interactions in living cells.
Supporting Evidence
- The split-TEV system allows for the monitoring of protein interactions in living cells.
- Phosphorylation of Bad at Ser136 is crucial for its interaction with 14-3-3 proteins.
- Split-TEV assays can be adapted for high-throughput screening of pharmacological substances.
Takeaway
This study shows a new way to see how proteins interact in cells when they are turned on by signals, which can help us understand diseases better.
Methodology
The study adapted the split-TEV system to quantify phosphorylation-dependent protein-protein interactions using genetically encoded reporters in NIH-3T3 fibroblasts and primary cultured neurons.
Limitations
The transfection efficiencies in primary cultured neurons were low, which may affect the robustness of the results.
Participant Demographics
The study involved NIH-3T3 fibroblasts and primary cultured neurons.
Digital Object Identifier (DOI)
Want to read the original?
Access the complete publication on the publisher's website