Peptide Assembly Directed and Quantified Using Megadalton DNA Nanostructures.

Area of Science:

• Biomolecular Engineering
• Supramolecular Chemistry
• Nanotechnology

Background:

• Nature utilizes co-assembly of biomolecules for functional structures.
• Quantifying multivalent interactions, especially low-affinity ones, remains challenging.

Purpose of the Study:

• To demonstrate DNA nanostructures as a platform for templating peptide interactions.
• To enable quantification of multivalent peptide-peptide interactions.
• To program spatial organization of synthetic biomolecular assemblies.

Main Methods:

• Design of peptide-oligonucleotide conjugates with de novo coiled-coil peptides.
• Integration of conjugates into DNA origami nanostructures.
• Quantification of nanostructure association via electron microscopy.
• Estimation of dissociation constants from binding data.

Main Results:

• DNA nanostructures successfully templated peptide-peptide interactions.
• Dissociation constants for single and multivalent peptide interactions were measured.
• Results align with solution-phase measurements and reveal polyvalency effects.
• Access to previously unquantifiable low-affinity interactions was achieved.

Conclusions:

• DNA nanostructures serve as effective scaffolds for studying peptide interactions.
• This approach facilitates the measurement of weak binding affinities and polyvalency.
• Programmable hybrid biomolecular assemblies can be constructed for advanced applications.