期刊
NUCLEIC ACIDS RESEARCH
卷 47, 期 11, 页码 5963-5972出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkz336
关键词
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资金
- Hubei Province [2016CFA002]
- National Natural Sciences Foundation of China [21190041, 21735007, 21605157, 21874149]
- Ministry of Science and Technology of China [2017YFA0505400]
- K.C. Wong Education Foundation
- National Natural Sciences Foundation of China
SELEX is the cornerstone for aptamer research with broad applications in biosensors and medicine. To improve the affinity of selected aptamers, we propose a structure-guided post-SELEX approach, an optimizationmethod based on the precise secondary structure of the aptamer-ligand complex. We demonstrate this approach using the Ochratoxin A (OTA) aptamer. Guided by the structure, we designed a new aptamer whose affinity is improved by more than 50-fold. We also determined the high-resolution NMR structure of the new aptamer-OTA complex and elucidated the discriminatory recognition mechanism of one atomic difference between two analogs, OTA and OTB. The aptamer forms an unusual hairpin structure containing an intramolecular triple helix, which is not seen in the previously determined aptamer complex. The pi-pi stacking, the hydrophobic interaction, hydrogen bonds and halogen bonds between OTA and the aptamer contribute to the recognition of OTA, and the halogen bonds play an important role in discriminating between OTA and OTB. Our results demonstrate that the structure-guided post-SELEX approach improves aptamers affinity. An improved OTA biosensor system might be developed using this new strategy.
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