Engineering a light-controlled F1 ATPase using structure-based protein design

Area of Science:

• Biochemistry
• Molecular Biology
• Nanotechnology

Background:

• ATP synthase's F1 sub-complex functions as a highly efficient biological nanomotor.
• This nanomotor converts chemical energy from ATP hydrolysis into mechanical work.
• Understanding its mechanics is crucial for bio-inspired machine design.

Purpose of the Study:

• To probe the mechanics of the F1 ATPase nanomotor.
• To engineer a synthetic ATPase with light-controlled activity.
• To explore structure-based protein design for dynamic constraint.

Main Methods:

• Employed a structure-based protein design approach to re-engineer the E. coli F1 ATPase active site.
• Incorporated a site-specific, photoswitchable crosslinker.
• Modulated inter-atomic distances between α and β subunits using light of different wavelengths.

Main Results:

• Crosslinking reduced ATP hydrolysis activity in four engineered designs.
• One design exhibited reversibly modulated ATPase activity upon illumination with near-UV and blue light.
• Demonstrated light-induced dynamic constraint on the nanomotor's subunits.

Conclusions:

• The study presents a novel method for creating light-controllable biological nanomachines.
• This work is a foundational step towards designing light-controlled nanomachines using biological components.
• Highlights the potential of protein engineering for creating synthetic molecular machines.