TY - JOUR
T1 - Folding Topology of a Short Coiled-Coil Peptide Structure Templated by an Oligonucleotide Triplex
AU - Lou, Chenguang
AU - Christensen, Niels Johan
AU - Martos Maldonado, Manuel Cristo
AU - Midtgaard, Søren Roi
AU - Ejlersen, Maria
AU - Thulstrup, Peter Waaben
AU - Sørensen, Kasper Kildegaard
AU - Jensen, Knud Jørgen
AU - Wengel, Jesper
PY - 2017/7/12
Y1 - 2017/7/12
N2 - The rational design of a well-defined protein-like tertiary structure formed by small peptide building blocks is still a formidable challenge. By using peptide-oligonucleotide conjugates (POC) as building blocks, we present the self-assembly of miniature coiled-coil α-helical peptides guided by oligonucleotide duplex and triplex formation. POC synthesis was achieved by copper-free alkyne-azide cycloaddition between three oligonucleotides and a 23-mer peptide, which by itself exhibited multiple oligomeric states in solution. The oligonucleotide domain was designed to furnish a stable parallel triplex under physiological pH, and to be capable of templating the three peptide sequences to constitute a small coiled-coil motif displaying remarkable α-helicity. The formed trimeric complex was characterized by ultraviolet thermal denaturation, gel electrophoresis, circular dichroism (CD) spectroscopy, small-angle X-ray scattering (SAXS), and molecular modeling. Stabilizing cooperativity was observed between the trimeric peptide and the oligonucleotide triplex domains, and the overall molecular size (ca. 12nm) in solution was revealed to be independent of concentration. The topological folding of the peptide moiety differed strongly from those of the individual POC strands and the unconjugated peptide, exclusively adopting the designed triple helical structure.
AB - The rational design of a well-defined protein-like tertiary structure formed by small peptide building blocks is still a formidable challenge. By using peptide-oligonucleotide conjugates (POC) as building blocks, we present the self-assembly of miniature coiled-coil α-helical peptides guided by oligonucleotide duplex and triplex formation. POC synthesis was achieved by copper-free alkyne-azide cycloaddition between three oligonucleotides and a 23-mer peptide, which by itself exhibited multiple oligomeric states in solution. The oligonucleotide domain was designed to furnish a stable parallel triplex under physiological pH, and to be capable of templating the three peptide sequences to constitute a small coiled-coil motif displaying remarkable α-helicity. The formed trimeric complex was characterized by ultraviolet thermal denaturation, gel electrophoresis, circular dichroism (CD) spectroscopy, small-angle X-ray scattering (SAXS), and molecular modeling. Stabilizing cooperativity was observed between the trimeric peptide and the oligonucleotide triplex domains, and the overall molecular size (ca. 12nm) in solution was revealed to be independent of concentration. The topological folding of the peptide moiety differed strongly from those of the individual POC strands and the unconjugated peptide, exclusively adopting the designed triple helical structure.
KW - Circular dichroism
KW - Click chemistry
KW - Molecular modeling
KW - Oligonucleotides
KW - Peptides
U2 - 10.1002/chem.201700971
DO - 10.1002/chem.201700971
M3 - Journal article
C2 - 28383784
AN - SCOPUS:85019120398
SN - 0947-6539
VL - 23
SP - 9297
EP - 9305
JO - Chemistry: A European Journal
JF - Chemistry: A European Journal
IS - 39
ER -