Covalently-assembled single-chain protein nanostructures with ultra-high stability

Area of Science:

• Biochemistry
• Materials Science
• Nanotechnology

Background:

• Protein nanostructures are crucial for applications like catalysis and drug delivery.
• Current methods primarily use non-covalent interactions for assembly.
• Covalent linking of proteins remains an underexplored strategy for nanostructure construction.

Purpose of the Study:

• To develop a novel method for creating protein nanostructures using covalent ligation.
• To synthesize single-chain protein nanostructures with defined geometries (triangular and square).
• To investigate the stability and applicability of these novel protein assemblies.

Main Methods:

• Utilized site-specific ligation via split intein chemistry.
• Employed multiple copies of a three-helix bundle protein.
• Performed coarse-grained simulations to analyze nanostructure flexibility.

Main Results:

• Successfully synthesized single-chain protein nanostructures with triangular and square shapes.
• Confirmed nanostructure flexibility through simulations, optimizing triangular structures for regularity.
• Demonstrated ultra-high thermostability and resistance to denaturation.
• Showcased applicability as scaffolds for nanoscale material assembly.

Conclusions:

• Site-specific covalent ligation is an effective strategy for assembling individually folded proteins into single-chain nanostructures.
• These protein nanostructures possess bespoke architectures and exceptional stability.
• The developed method opens avenues for advanced applications in catalysis, sensing, and materials science.