Structural and Binding Effects of Chemical Modifications on Thrombin Binding Aptamer (TBA)
| Autor: | Yicheng Huang, Jia Sheng, Alan A. Chen, Sweta Vangaveti, Vibhav A. Valsangkar, Goh Woon Lee, Walid M. Fahssi, Waqas S. Awan |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Circular dichroism Aptamer Oligonucleotides Pharmaceutical Science Organic chemistry Molecular Dynamics Simulation 030204 cardiovascular system & hematology Oligonucleotide synthesis Article Analytical Chemistry dendrimers 03 medical and health sciences 0302 clinical medicine Thrombin QD241-441 Dendrimer Drug Discovery medicine Humans Physical and Theoretical Chemistry Surface plasmon resonance Binding site Blood Coagulation trebler Binding Sites Base Sequence Chemistry Anticoagulants Aptamers Nucleotide Combinatorial chemistry G-Quadruplexes Dissociation constant thrombin binding aptamer (TBA) Kinetics 030104 developmental biology Chemistry (miscellaneous) NHS-carboxy T Nucleic Acid Conformation Thermodynamics Molecular Medicine Protein Binding medicine.drug |
| Zdroj: | Molecules, Vol 26, Iss 4620, p 4620 (2021) Molecules Volume 26 Issue 15 |
| ISSN: | 1420-3049 |
| Popis: | The thrombin binding aptamer (TBA) is a promising nucleic acid-based anticoagulant. We studied the effects of chemical modifications, such as dendrimer Trebler and NHS carboxy group, on TBA with respect to its structures and thrombin binding affinity. The two dendrimer modifications were incorporated into the TBA at the 5′ end and the NHS carboxy group was added into the thymine residues in the thrombin binding site of the TBA G-quadruplex (at T4, T13 and both T4/T13) using solid phase oligonucleotide synthesis. Circular dichroism (CD) spectroscopy confirmed that all of these modified TBA variants fold into a stable G-quadruplex. The binding affinity of TBA variants with thrombin was measured by surface plasmon resonance (SPR). The binding patterns and equilibrium dissociation constants (KD) of the modified TBAs are very similar to that of the native TBA. Molecular dynamics simulations studies indicate that the additional interactions or stability enhancement introduced by the modifications are minimized either by the disruption of TBA–thrombin interactions or destabilization elsewhere in the aptamer, providing a rational explanation for our experimental data. Overall, this study identifies potential positions on the TBA that can be modified without adversely affecting its structure and thrombin binding preference, which could be useful in the design and development of more functional TBA analogues. |
| Databáze: | OpenAIRE |
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