title: Study of the interaction between the G-quadruplex-forming thrombin-binding aptamer and the porphyrin 5,10,15,20-tetrakis-(N-methyl-4-pyridyl)-21,23H-porphyrin tetratosylate.
abstract: The G-quadruplex DNA structure has been suggested to be a potential target for anticancer therapies. Therefore, there is increasing interest in the development of drugs that could modulate the stability of G-quadruplex structures. In the current work, the interaction between the thrombin-binding aptamer (TBA, 5'-GGT TGG TGT GGT TGG-3'), which can form an intramolecular G-quadruplex structure, and the porphyrin 5,10,15,20-tetrakis-(N-methyl-4-pyridyl)-21,23H-porphyrin tetratosylate (TmPyP4) was studied. The application of a high-performance liquid chromatography-photodiode array (HPLC-PDA) detector-based method to study this kind of interaction was tested. Molecular absorption data recorded along the chromatographic runs were analyzed by means of multivariate data analysis methods. Moreover, biospecific interaction analysis (BIA) by surface plasmon resonance (SPR) and melting and mole ratio experiments monitored by UV-visible molecular absorption and circular dichroism spectroscopies, were applied to confirm and expand the chromatographic studies. The results showed the formation of an interaction complex with a stoichiometry 1:1 (TmPyP4/TBA) and logarithm of the equilibrium constant equal to 5.7+/-0.2. Melting and circular dichroism data reflected that the initial G-quadruplex structure of TBA is stabilized in the interaction complex, being slightly distorted by the presence of the ligand.
authors.affiliation: Department of Analytical Chemistry, Universitat de Barcelona, Barcelona E-08028, Spain.
authors.forename: Miquel
authors.initials: M
authors.lastname: del Toro
authors.forename: Raimundo
authors.initials: R
authors.lastname: Gargallo
authors.forename: Ramon
authors.initials: R
authors.lastname: Eritja
authors.forename: Joaquim
authors.initials: J
authors.lastname: Jaumot
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D054856:G-Quadruplexes; D015394:Molecular Structure; D011166:Porphyrins / Q000737:chemistry*; D020349:Surface Plasmon Resonance
chemical_list: D052157:Aptamers, Nucleotide; D011166:Porphyrins; C081676:thrombin aptamer; C021096:tetra(4-N-methylpyridyl)porphine