Article ; Online: Thermodynamic analysis of cooperative ligand binding by the ATP-binding DNA aptamer indicates a population-shift binding mechanism.
2020 Volume 10, Issue 1, Page(s) 18944
Abstract: The ATP-binding DNA aptamer is often used as a model system for developing new aptamer-based biosensor methods. This aptamer follows a structure-switching binding mechanism and is unusual in that it binds two copies of its ligand. We have used isothermal ...
| Abstract | The ATP-binding DNA aptamer is often used as a model system for developing new aptamer-based biosensor methods. This aptamer follows a structure-switching binding mechanism and is unusual in that it binds two copies of its ligand. We have used isothermal titration calorimetry methods to study the binding of ATP, ADP, AMP and adenosine to the ATP-binding aptamer. Using both individual and global fitting methods, we show that this aptamer follows a positive cooperative binding mechanism. We have determined the binding affinity and thermodynamics for both ligand-binding sites. By separating the ligand-binding sites by an additional four base pairs, we engineered a variant of this aptamer that binds two adenosine ligands in an independent manner. Together with NMR and thermal stability experiments, these data indicate that the ATP-binding DNA aptamer follows a population-shift binding mechanism that is the source of the positive binding cooperativity by the aptamer. |
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| MeSH term(s) | Adenosine Triphosphate/metabolism ; Aptamers, Nucleotide/chemistry ; Aptamers, Nucleotide/metabolism ; Binding Sites ; Calorimetry ; Magnetic Resonance Spectroscopy ; Thermodynamics |
| Chemical Substances | Aptamers, Nucleotide ; Adenosine Triphosphate (8L70Q75FXE) |
| Language | English |
| Publishing date | 2020-11-03 |
| Publishing country | England |
| Document type | Journal Article ; Research Support, Non-U.S. Gov't |
| ZDB-ID | 2615211-3 |
| ISSN | 2045-2322 ; 2045-2322 |
| ISSN (online) | 2045-2322 |
| ISSN | 2045-2322 |
| DOI | 10.1038/s41598-020-76002-8 |
| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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