On the magnitude of the chelate effect for the recognition of proteins by pharmacophores scaffolded by self-assembling oligonucleotides.
The simultaneous interaction of the binding moieties of a bidentate ligand on adjacent epitopes of a target protein represents an attractive avenue for the discovery of specific, high-affinity binders. We used short DNA fragments in heteroduplex format to scaffold pairs of binding molecules with defined spatial arrangements. Iminobiotin derivates were coupled either via bifunctional linkers or by using various oligonucleotides, thus allowing monovalent or bivalent binding to streptavidin. We determined the binding affinities of the synthesized constructs in solution. We also investigated the efficiency of recovery of superior bidentate ligands in affinity capture experiments, by using both radioactive counts and DNA microarrays as readouts. This analysis confirmed the suitability of the DNA heteroduplex as a scaffold for the identification of synergistic pairs of binding moieties, capable of a high-affinity interaction with protein targets by virtue of the chelate effect.
Melkko
Samu
S
ETH Zürich, Institute of Pharmaceutical Sciences, Switzerland.
Dumelin
Christoph E
CE
Scheuermann
Jörg
J
Neri
Dario
D
D001483:Base Sequence; D002151:Calorimetry; D002614:Chelating Agents / Q000737:chemistry*; D017931:DNA Primers; D020411:Oligonucleotide Array Sequence Analysis; D009841:Oligonucleotides / Q000737:chemistry*; D004364:Pharmaceutical Preparations / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*
D002614:Chelating Agents; D017931:DNA Primers; D009841:Oligonucleotides; D004364:Pharmaceutical Preparations; D011506:Proteins