Probing the Dynamic Interaction between Damaged DNA and a Cellular Responsive Protein Using a Piezoelectric Mass...

Area of Science:

• Biomolecular Interactions
• Biosensing Technology
• Drug Discovery

Background:

• Understanding DNA damage and repair is crucial for cancer therapy.
• High-mobility-group box 1 (HMGB1) is a protein involved in DNA damage response.
• Cisplatin is a widely used DNA-damaging anticancer drug.

Purpose of the Study:

• To develop a sensitive biosensor for real-time monitoring of interactions between cisplatin-damaged DNA (cisPt-DNA) and HMGB1.
• To investigate the kinetics and affinity of the cisPt-DNA and HMGB1a interaction.
• To provide a platform for evaluating anticancer drugs.

Main Methods:

• Integration of flow injection analysis (FIA) with quartz crystal microbalance (QCM).
• Fabrication of a specific sensing interface using cisPt-DNA as the recognition element.
• Utilizing a hybrid self-assembled monolayer to minimize nonspecific adsorption.
• Molecular docking to simulate the complex formation.

Main Results:

• The biosensor successfully detected and quantified the interaction between cisPt-DNA and HMGB1a in real-time.
• Kinetic parameters (k_ass, k_diss) and dissociation constant (K_D) indicated rapid recognition and tight binding.
• Molecular docking provided insights into the binding mechanism.

Conclusions:

• The developed continuous-flow QCM biosensor is effective for studying drug-damaged DNA and protein interactions.
• This system offers a potential platform for rapid screening and evaluation of metal-based anticancer drugs.
• The findings contribute to understanding DNA repair resistance mechanisms.