A structure-activity relationship of a thrombin-binding aptamer containing LNA in novel sites |
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| Institution: | 1. Laboratory of Analytical Chemistry, Department of Chemistry, University of Helsinki, PO. Box 55, FI-00014, Finland;2. Wihuri Research Institute, Haartmaninkatu 8, FI-00290 Helsinki, Finland;3. National Institute for Health and Welfare, Genomics and Biomarkers Unit, Biomedicum, FI-00290, Helsinki, Finland;4. Laboratory of Inorganic Chemistry, Department of Chemistry, University of Helsinki, PO. Box 55, FI-00014, Finland;1. Biomolecular Nanoscale Engineering Center, Department of Physics, Chemistry and Pharmacy, University of Southern Denmark, Campusvej 55, 5230 Odense M, Denmark;2. Department of Chemistry and Biochemistry, University of Colorado, Boulder, CO 80303, USA;3. Department of Chemistry, Faculty of Science, Damietta University, New Damietta 34517, Egypt;1. CICS-UBI – Centro de Investigação em Ciências da Saúde, Universidade da Beira Interior, Av. Infante D. Henrique, 6200-506 Covilhã, Portugal;2. Fluorescence Imaging Group, Departamento de Fisiología – Facultad de Medicina, Avda. Arzobispo Morcillo 2, Universidad Autónoma de Madrid, Madrid 28029, Spain;3. Nanobiology Group, Instituto Ramón y Cajal de Investigación Sanitaria, IRYCIS, Ctra. Colmenar km. 9.100, Madrid 28034, Spain;4. Fluorescence Imaging Group, Departamento de Física de Materiales, Facultad de Ciencias, Universidad Autónoma de Madrid, C/Francisco Tomás y Valiente 7, Madrid 28049, Spain;1. Furtwangen University, Institute of Precision Medicine, Jakob-Kienzle-Straße 17, 78054, Villingen-Schwenningen, Germany;2. Department of Pharmaceutical and Medicinal Chemistry, Institute of Pharmaceutical Sciences, Eberhard Karls Universität Tübingen, Auf der Morgenstelle 8, 72076, Tübingen, Germany;3. Fraunhofer Institute IZI, Leipzig, EXIM Department, Schillingallee 68, D-18057, Rostock, Germany |
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| Abstract: | In this report, structural characterization, aptamer stability and thrombin of a new modified thrombin-ligand complex binding aptamer (TBA) containing anti-guanine bases and a loop position locked nucleic acid (LNA) are presented. NMR, circular dichroic spectroscopy and molecular modeling were used to characterize the three-dimensional structure of two G-quadruplexes. LNA-modification of the anti-guanosines yields G-quadruplexes that show affinity and inhibitory activity toward thrombin, whereas LNA-modification of a thymine nucleotide in the TGT loop increases the thermal stability of TBA. As assessed by denatured PAGE electrophoresis, all modified aptamers display an increase in environmental stability. The prothrombin time assay and fibrinogen assay showed that the aptamers still had good inhibitory activity, and 15 of them had the longest PT time. Therefore, the LNA modification is well suited to improve the physicochemical and biological properties of the native thrombin-binding aptamer. |
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| Keywords: | Aptamer Locked nucleic acid Oligonucleotides TBA Anticoagulant activity |
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