文献类型：期刊文献

英文题名：Monitoring of an ATP-Binding Aptamer and its Conformational Changes Using an α-Hemolysin Nanopore

作者：Ying, Yi-Lun[1,2];Wang, Hai-Yan[1,2];Sutherland, Todd C.[3];Long, Yi-Tao[1,2]

机构：[1]E China Univ Sci & Technol, Key Lab Adv Mat, Shanghai 200237, Peoples R China;[2]E China Univ Sci & Technol, Dept Chem, Shanghai 200237, Peoples R China;[3]Univ Calgary, Dept Chem, Calgary, AB T2N 1N4, Canada

年份：2011

卷号：7

期号：1

起止页码：87

外文期刊名：SMALL

收录：;EI(收录号：20110113550095);WOS:【SCI-EXPANDED(收录号:WOS:000285794100010)】；

基金：This work was supported by the National Natural Science Foundation of China (Grant No. 20875030) and the Shuguang Project of Shanghai (Grant No. 07SG36). We would like to thank Professor Hong-Yuan Chen at Nanjing University for useful discussions.

语种：英文

外文关键词：Conformations - Nanopores - Adenosinetriphosphate - Biosensors - Blood - Oligonucleotides

摘要：An aptamer is a specific oligonucleotide sequence that spontaneously forms a secondary structure capable of selectively binding an analyte. An aptamer's conformation is the key to specific binding of a target molecule, even in the case of very closely related targets. Nanopores are a sensitive tool for the single-molecule analysis of DNA, peptides, and proteins transporting through the pore. Herein, a single alpha-hemolysin natural nanopore is utilized to sense the conformational changes of an adenosine 5'-triphosphate (ATP)-binding aptamer (ABA). The known DNA sequence of the ABA is used as a model to develop real-time monitoring of molecular conformational changes that occur by binding targets. The native, folded ABA structure has a nanopore unfolding time of 4.17 ms, compared with 0.29 ms for the ABA:ATP complex. A complementary 14-mer strand, which binds the ABA sequence in the key nucleic acids responsible for folding, forms linear duplex DNA, resulting in a nanopore transit time of 0.50 ms and a higher capture probability than that of the folded ABA oligomer. Competition assays between the ABA: ATP and ABA: reporter complexes are carried out, and the results suggest that the ABA: ATP complex is formed preferentially. The nanopore allows for the detection of an ABA in its folded, ATP-bound, and linear conformations.