Fluorogenic resolution of ligand binding by a nucleic acid aptamer.
An ATP-binding nucleic acid aptamer was converted into a solution-phase fluorescence-detected sensor by incorporating a 2'-amine group in a structure that is sensitive to a ligand-induced conformational change in the aptamer. The 2'-amine substitution in the free aptamer reacts efficiently with fluorescamine to yield a fluorescent product detectable by energy transfer to a tethered fluor emitting at 615 nm. In the presence of ATP, the fluorogenic reaction is substantially reduced, both under simplified conditions and in a urine background. Fluorogenic chemistry represents a potentially general approach for creating sensors from aptamers that bind their ligands via induced fit.
Merino
Edward J
EJ
Department of Chemistry, University of North Carolina, Chapel Hill, North Carolina 27599-3290, USA.
Weeks
Kevin M
KM
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry; D015374:Biosensing Techniques / Q000379:methods; D004247:DNA / Q000737:chemistry; D005450:Fluorescamine / Q000737:chemistry; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry; D006801:Humans; D008024:Ligands; D012313:RNA / Q000737:chemistry
D005456:Fluorescent Dyes; D008024:Ligands; D005450:Fluorescamine; D012313:RNA; D000255:Adenosine Triphosphate; D004247:DNA