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- Title
Novel application of fluorescence coupled capillary electrophoresis to resolve the interaction between the G-quadruplex aptamer and thrombin.
- Authors
Wang, Jianhao; Gu, Yaqin; Liu, Li; Wang, Cheli; Wang, Jianpeng; Ding, Shumin; Li, Jinping; Qiu, Lin; Jiang, Pengju
- Abstract
The dynamic binding status between the thrombin and its G-quadruplex aptamers and the stability of its interaction partners were probed using our previously established fluorescence-coupled capillary electrophoresis method. A 29-nucleic acid thrombin binding aptamer was chosen as a model to study its binding affinity with the thrombin ligand. First, the effects of the cations on the formation of G-quadruplex from unstructured 29-nucleic acid thrombin binding aptamer were examined. Second, the rapid binding kinetics between the thrombin and 6-carboxyfluorescein labeled G-quadruplex aptamer was measured. Third, the stability of G-quadruplex aptamer-thrombin complex was also examined in the presence of the interfering species. Remarkably, it was found that the complementary strand of 29-nucleic acid thrombin binding aptamer could compete with G-quadruplex aptamer and thus disassociated the G-quadruplex structure into an unstructured aptamer. These data suggest that our in-house established fluorescence-coupled capillary electrophoresis assay could be applied to binding studies of the G-quadruplex aptamers, thrombin, and their ligands, while overcoming the complicated and costly approaches currently available.
- Subjects
APTAMERS; THROMBIN; CAPILLARY electrophoresis; QUADRUPLEX nucleic acids; LIGANDS (Biochemistry)
- Publication
Journal of Separation Science, 2017, Vol 40, Issue 15, p3161
- ISSN
1615-9306
- Publication type
Article
- DOI
10.1002/jssc.201700456