Capillary gel electrophoresis analysis of G-quartet forming oligonucleotides used in DNA-protein interaction studies.
DNA-protein binding is among the most frequently studied biomolecular interactions with high importance in modern systems biology research. One interesting aspect of this rapidly developing field is the affinity capture of proteins by G-quartet forming oligonucleotides also referred to as aptamers. G-quartets are structural motifs formed by guanine-rich sequences commonly occurring in the human genome. In this paper, we describe a capillary gel electrophoresis based method to validate G-quartet formation of in-house designed oligonucleotides and discuss the effect of monovalent cation concentration on the development of this structure. The relevant aptamer was then bound to magnetic beads to form an affinity capture surface for target proteins, which were then analyzed by matrix-assisted laser desorption/ionization mass spectrometry.
Szilagyi
Agnes
A
Horváth Laboratory of Bioseparation Sciences, Institute of Analytical Chemistry and Radiochemistry, Leopold-Franzens University, Innrain 66, A-6020 Innsbruck, Austria.
Bonn
Günther K
GK
Guttman
András
A
D052157:Aptamers, Nucleotide; D001483:Base Sequence; D004247:DNA / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D052157:Aptamers, Nucleotide; D011506:Proteins; D004247:DNA