Peptide Assembly Directed and Quantified Using Megadalton DNA Nanostructures

Juan Jin, Emily Baker, Christopher Wood, Jonathan Bath, Dek Woolfson, Andrew Tuberfield

Research output: Contribution to journalArticle (Academic Journal)peer-review

12 Citations (Scopus)
240 Downloads (Pure)

Abstract

In nature, co-assembly of polypeptides, nucleic acids, and polysaccharides is used to create functional supramolecular structures. Here, we show that DNA nanostructures can be used to template interactions between peptides and to enable the quantification of multivalent interactions that would otherwise not be observable. Our functional building blocks are peptide–oligonucleotide conjugates comprising de novo designed dimeric coiled-coil peptides covalently linked to oligonucleotide tags. These conjugates are incorporated in megadalton DNA origami nanostructures and direct nanostructure association through peptide–peptide interactions. Free and bound nanostructures can be counted directly from electron micrographs, allowing estimation of the dissociation constants of the peptides linking them. Results for a single peptide–peptide interaction are consistent with the measured solution-phase free energy; DNA nanostructures displaying multiple peptides allow the effects of polyvalency to be probed. This use of DNA nanostructures as identifiers allows the binding strengths of homo- and heterodimeric peptide combinations to be measured in a single experiment and gives access to dissociation constants that are too low to be quantified by conventional techniques. The work also demonstrates that hybrid biomolecules can be programmed to achieve spatial organization of complex synthetic biomolecular assemblies.
Original languageEnglish
Number of pages9
JournalACS Nano
Early online date8 Aug 2019
DOIs
Publication statusE-pub ahead of print - 8 Aug 2019

Structured keywords

  • BrisSynBio
  • Bristol BioDesign Institute

Keywords

  • self-assembly
  • DNA nanostructure
  • peptide
  • polyvalent binding
  • heterodimeric coiled coil
  • peptide−oligonucleotide conjugate
  • Synthetic biology

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