Related Experiment Video
Updated: Apr 7, 2026
14:12
Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells
Published on: December 11, 2021
6.2K
In Situ Study of p53, MDM2, and MDMX Protein Interaction in Living Cells Using Fluorescence Triple Correlation
Xinwei Lu1, Kun Zang1, Chaoliang Diao1
1School of Chemistry and Chemical Engineering, Shanghai Jiao Tong University 800 Dongchuan Road, Shanghai 200240, P. R. China.
Analytical Chemistry
|April 6, 2026
Summary
Researchers developed a new method to observe the p53-MDM2-MDMX protein complex in living cells. This technique reveals how MDMX structure affects complex stability and evaluates p53 inhibitor efficacy in real-time.
Area of Science:
- Molecular Biology
- Cell Biology
- Biophysics
Background:
- The tumor suppressor protein p53 is vital for cellular regulation, with its activity controlled by MDM2 and MDMX.
- Studying p53, MDM2, and MDMX interactions in living cells (in situ) is challenging.
Purpose of the Study:
- To develop a novel method for investigating p53, MDM2, and MDMX interactions in situ.
- To observe and quantify the p53-MDM2-MDMX ternary complex formation in living cells.
- To assess the impact of MDMX structure on complex stability and evaluate p53 inhibitor efficacy.
Main Methods:
- Combined fluorescence triple correlation spectroscopy (FTCS) with protein fusion labeling.
- Fluorescently labeled p53, MDM2, and MDMX using gene engineering.
- Quantified ternary complex concentration and dissociation kinetics.
Main Results:
- Achieved the first in situ observation of the p53-MDM2-MDMX ternary complex in living cells.
- Demonstrated that the MDMX RING domain significantly influences ternary complex stability.
- Developed a method to evaluate p53 inhibitor efficacy, showing RO-5963 rapidly disrupts complex stability.
Conclusions:
- The developed FTCS method enables real-time analysis of protein complex dynamics in living cells.
- Understanding ternary complex formation and dissociation is crucial for cancer research.
- This approach provides significant insights into tumorigenesis mechanisms.
