Dynamics of an anti-VEGF DNA aptamer: a single-molecule study.
Single-molecule fluorescence resonance energy transfer (SMFRET) was used to study the interaction of a 25-nucleotide (nt) DNA aptamer with its binding target, vascular endothelial growth factor (VEGF). Conformational dynamics of the aptamer were studied in the absence of VEGF in order to characterize fluctuations in the unbound nucleic acid. SMFRET efficiency distributions showed that, while the aptamer favors a base-paired conformation, there are frequent conversions to higher energy conformations. Conversions to higher energy structures were also demonstrated to be dependent on the concentration of Mg2+-counterion by an overall broadening of the SMFRET efficiency distribution at lower Mg2+ concentration. Introduction of VEGF caused a distinct increase in the frequency of lower SMFRET efficiencies, indicating that favorable interaction of the DNA aptamer with its VEGF target directs aptamer structure towards a more open conformation.
Department of Chemistry, 136 Fleming, University of Houston, Houston, TX 77204-5003, USA.
J
J
Nick Taylor
Qusai
Q
Darugar
Katerina
K
Kourentzi
Richard C
RC
Willson
Christy F
CF
Landes
D052157:Aptamers, Nucleotide / Q000737:chemistry*; D001483:Base Sequence; D004247:DNA / Q000737:chemistry*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D008274:Magnesium / Q000737:chemistry; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D042461:Vascular Endothelial Growth Factor A / Q000737:chemistry*
D052157:Aptamers, Nucleotide; D042461:Vascular Endothelial Growth Factor A; D004247:DNA; D008274:Magnesium