Peptide-oligonucleotide conjugates as nanoscale building blocks for assembly of an artificial three-helix protein mimic

Chenguang Lou, Manuel C Martos-Maldonado, Charlotte S Madsen, Rasmus P Thomsen, Søren Roi Midtgaard, Niels Johan Christensen, Jørgen Kjems, Peter W Thulstrup, Jesper Wengel, Knud J Jensen

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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.

Original languageEnglish
Article number12294
JournalNature Communications
Volume7
Pages (from-to)1-9
ISSN2041-1723
DOIs
Publication statusPublished - 2016

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oligonucleotides
Oligonucleotides
helices
peptides
assembly
proteins
Peptides
Self assembly
Circular dichroism spectroscopy
self assembly
Circular Dichroism
Proteins
assemblies
dichroism
coils
Transmission Electron Microscopy
Freezing
electrophoresis
trimers
Electrophoresis

Keywords

  • Journal Article

Cite this

Lou, Chenguang ; Martos-Maldonado, Manuel C ; Madsen, Charlotte S ; Thomsen, Rasmus P ; Midtgaard, Søren Roi ; Christensen, Niels Johan ; Kjems, Jørgen ; Thulstrup, Peter W ; Wengel, Jesper ; Jensen, Knud J. / Peptide-oligonucleotide conjugates as nanoscale building blocks for assembly of an artificial three-helix protein mimic. In: Nature Communications. 2016 ; Vol. 7. pp. 1-9.
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Lou, C, Martos-Maldonado, MC, Madsen, CS, Thomsen, RP, Midtgaard, SR, Christensen, NJ, Kjems, J, Thulstrup, PW, Wengel, J & Jensen, KJ 2016, 'Peptide-oligonucleotide conjugates as nanoscale building blocks for assembly of an artificial three-helix protein mimic', Nature Communications, vol. 7, 12294, pp. 1-9. https://doi.org/10.1038/ncomms12294

Peptide-oligonucleotide conjugates as nanoscale building blocks for assembly of an artificial three-helix protein mimic. / Lou, Chenguang; Martos-Maldonado, Manuel C; Madsen, Charlotte S; Thomsen, Rasmus P; Midtgaard, Søren Roi; Christensen, Niels Johan; Kjems, Jørgen; Thulstrup, Peter W; Wengel, Jesper; Jensen, Knud J.

In: Nature Communications, Vol. 7, 12294, 2016, p. 1-9.

Research output: Contribution to journalJournal articleResearchpeer-review

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