Analysis of the interaction between the cocaine-binding aptamer and its ligands using fluorescence spectroscopy

被引:31
|
作者
Shoara, Aron A. [1 ,2 ]
Slavkovic, Sladjana [1 ,2 ]
Donaldson, Logan W. [1 ,2 ]
Johnson, Philip E. [1 ,2 ]
机构
[1] York Univ, Dept Chem, Toronto, ON M3J 1P3, Canada
[2] York Univ, Ctr Res Biomol Interact, Toronto, ON M3J 1P3, Canada
关键词
fluorescence spectroscopy; fluorescence quenching; aptamers; small molecule - DNA interactions; LABEL-FREE DETECTION; DNA; SENSORS; CALORIMETRY; MECHANISM; SPECTRA; DESIGN; TIME;
D O I
10.1139/cjc-2017-0380
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
We used fluorescence spectroscopy to measure the binding affinity and provide new insights into the binding mechanism of cocaine and quinine with the cocaine-binding DNA aptamer. Using the intrinsic fluorescence of quinine and cocaine, we have observed quenching of ligand fluorescence upon binding of the aptamer. Quantification of this quenching provides an easy method to measure the binding constant using small amounts of sample. The observed quenching coupled with a red shift of the Stokes shift in the emission spectrum indicates that quinine and cocaine interact with the aptamer through stacking interactions.
引用
收藏
页码:1253 / 1260
页数:8
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