Mapping molecular binding by means of conformational dynamics measurements

Area of Science:

• Biochemistry
• Structural Biology
• Biophysics

Background:

• Protein-protein interactions are fundamental to biological processes.
• Understanding these interactions and their interfaces is crucial for deciphering biological function.
• Observing protein-protein interactions in their dynamic, native environment presents significant technical challenges.

Purpose of the Study:

• To develop and validate a novel method for real-time, in situ monitoring of protein-protein interaction dynamics.
• To advance technological platforms for drug discovery by enabling dynamic interaction studies.
• To investigate the conformational dynamics of specific protein-antibody interactions.

Main Methods:

• Integration of dual polarization interferometry (DPI) and quartz crystal microbalance with dissipation monitoring (QCM-D).
• Real-time, in situ monitoring of conformational changes during protein-protein binding events.
• Application of a chip-based assay for high-throughput screening.

Main Results:

• Successfully mapped the conformational dynamics of protein-protein interactions in real time.
• Demonstrated the method's efficacy in studying the interaction between TRIM21α protein and autoantibodies via a bridging bipolar mechanism.
• Validated the combined DPI-QCM-D approach for studying dynamic molecular interactions.

Conclusions:

• The developed chip-based method offers a significant technological advancement for studying protein-protein interaction dynamics.
• This approach facilitates miniaturized, high-throughput determination of interaction mechanisms.
• The findings support the application of this technique in drug discovery and fundamental biological research.