title: Peptide-oligonucleotide conjugates as nanoscale building blocks for assembly of an artificial three-helix protein mimic.
abstract: Peptide-based structures can be designed to yield artificial proteins with specific folding patterns and functions. Template-based assembly of peptide units is one design option, but the use of two orthogonal self-assembly principles, oligonucleotide triple helix and a coiled coil protein domain formation have never been realized for de novo protein design. Here, we show the applicability of peptide-oligonucleotide conjugates for self-assembly of higher-ordered protein-like structures. The resulting nano-assemblies were characterized by ultraviolet-melting, gel electrophoresis, circular dichroism (CD) spectroscopy, small-angle X-ray scattering and transmission electron microscopy. These studies revealed the formation of the desired triple helix and coiled coil domains at low concentrations, while a dimer of trimers was dominating at high concentration. CD spectroscopy showed an extraordinarily high degree of α-helicity for the peptide moieties in the assemblies. The results validate the use of orthogonal self-assembly principles as a paradigm for de novo protein design.
authors.affiliation: Department of Physics, Chemistry and Pharmacy, Biomolecular Nanoscale Engineering Center, University of Southern Denmark, Campusvej 55, Odense M 5230, Denmark.
authors.forename: Chenguang
authors.initials: C
authors.lastname: Lou
authors.affiliation: Department of Chemistry, Biomolecular Nanoscale Engineering Center, University of Copenhagen, Thorvaldsensvej 40, Frederiksberg 1871, Denmark.
authors.forename: Manuel C
authors.initials: MC
authors.lastname: Martos-Maldonado
authors.affiliation: Department of Chemistry, Biomolecular Nanoscale Engineering Center, University of Copenhagen, Thorvaldsensvej 40, Frederiksberg 1871, Denmark.
authors.forename: Charlotte S
authors.initials: CS
authors.lastname: Madsen
authors.affiliation: Biomolecular Nanoscale Engineering Center and Interdisciplinary Nanoscience Center (iNANO), University of Aarhus, Gustav Wieds Vej 14, Aarhus C 8000, Denmark.
authors.forename: Rasmus P
authors.initials: RP
authors.lastname: Thomsen
authors.affiliation: Niels Bohr Institute, University of Copenhagen, Universitetsparken 5, Copenhagen Ø 2100, Denmark.
authors.forename: Søren Roi
authors.initials: SR
authors.lastname: Midtgaard
authors.affiliation: Department of Chemistry, Biomolecular Nanoscale Engineering Center, University of Copenhagen, Thorvaldsensvej 40, Frederiksberg 1871, Denmark.
authors.forename: Niels Johan
authors.initials: NJ
authors.lastname: Christensen
authors.affiliation: Biomolecular Nanoscale Engineering Center and Interdisciplinary Nanoscience Center (iNANO), University of Aarhus, Gustav Wieds Vej 14, Aarhus C 8000, Denmark.
authors.forename: Jørgen
authors.initials: J
authors.lastname: Kjems
authors.affiliation: Department of Chemistry, University of Copenhagen, Universitetsparken 5, Copenhagen Ø 2100, Denmark.
authors.forename: Peter W
authors.identifier: 0000-0002-9241-4352
authors.initials: PW
authors.lastname: Thulstrup
authors.affiliation: Department of Physics, Chemistry and Pharmacy, Biomolecular Nanoscale Engineering Center, University of Southern Denmark, Campusvej 55, Odense M 5230, Denmark.
authors.forename: Jesper
authors.initials: J
authors.lastname: Wengel
authors.affiliation: Department of Chemistry, Biomolecular Nanoscale Engineering Center, University of Copenhagen, Thorvaldsensvej 40, Frederiksberg 1871, Denmark.
authors.forename: Knud J
authors.initials: KJ
authors.lastname: Jensen
mesh_terms: D000595:Amino Acid Sequence; D002942:Circular Dichroism; D008958:Models, Molecular; D053758:Nanoparticles / Q000737:chemistry*; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000737:chemistry*; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry*; D011489:Protein Denaturation; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry*; D053838:Scattering, Small Angle; D014466:Ultraviolet Rays; D014961:X-Ray Diffraction
chemical_list: D009841:Oligonucleotides; D010455:Peptides; D011506:Proteins