Directly watching biomolecules in action by high-speed atomic force microscopy

Area of Science:

• Biophysics
• Molecular Biology
• Microscopy

Background:

• Proteins function dynamically at the single-molecule level.
• Understanding protein function requires observing them in action.
• Existing methods may not capture rapid molecular events.

Purpose of the Study:

• To introduce and highlight the capabilities of high-speed atomic force microscopy (HS-AFM).
• To demonstrate HS-AFM's utility in visualizing dynamic protein processes.
• To showcase HS-AFM's potential for molecular manipulation and discovery.

Main Methods:

• Development and application of high-speed atomic force microscopy (HS-AFM).
• Imaging biological molecules at 10-16 frames/second without functional disturbance.
• Utilizing the cantilever tip for in-situ molecular manipulation and perturbation.

Main Results:

• Successful visualization of dynamic protein structures and processes (e.g., bacteriorhodopsin, myosin V, disordered proteins).
• Acquisition of molecular movies providing unique mechanistic insights.
• New discoveries through manipulation-based imaging of myosin V, peroxiredoxin, and microtubules.

Conclusions:

• HS-AFM is a powerful tool for observing protein dynamics in real-time.
• HS-AFM enables direct insights into molecular mechanisms through visualization and manipulation.
• Future applications will expand with HS-AFM integration with other advanced techniques.