A new method for the determination of the relative affinity of a ligand against various DNA sequences by electrospray ionization mass spectrometry. Application to a polyamide minor groove binder.
A new method for the determination of the relative affinity of a ligand against various dsDNA sequences is presented by using electrospray ionization time-of-flight mass spectrometry (ESI-QTOF) mass spectrometry. The principle is described here through the complexation of double-stranded DNA by a polyamide ligand including twelve N-methylpyrrole rings. However this method could be applied to other ligands especially when dissociation constants (Kd) are in nanomolar range. This method does not require knowing the ligand concentration accurately. It allows determination of the relative affinity of a ligand against various dsDNA sequences for 1 : 1 complex stoichiometries in a quick manner without labeling.
Buchmann
William
W
Université d'Evry val d'Essonne, Laboratoire Analyse et Modélisation pour la Biologie et l'Environnement, (CNRS UMR 8587/CEA), Bâtiment Maupertuis, Bd. François Mitterrand, 91025 Evry, France. william.buchmann@univ-evry.fr
Boutorine
Alexandre
A
Halby
Ludovic
L
Tortajada
Jeanine
J
De Pauw
Edwin
E
D001665:Binding Sites; D001667:Binding, Competitive; D004247:DNA / Q000737:chemistry*; D008024:Ligands; D008958:Models, Molecular; D009757:Nylons / Q000737:chemistry*; D009841:Oligonucleotides / Q000737:chemistry*; D011758:Pyrroles; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry
D008024:Ligands; D009757:Nylons; D009841:Oligonucleotides; D011758:Pyrroles; D004247:DNA; C096654:N-methylpyrrole