Selective recognition of parallel and anti-parallel thrombin-binding aptamer G-quadruplexes by different fluorescent dyes.
Selective recognition of parallel and anti-parallel thrombin-binding aptamer G-quadruplexes by different fluorescent dyes.
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不同荧光染料选择性识别平行和反平行凝血酶结合适体 G 四链体
DOI：
10.1093/nar/gku833
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发表时间：
2014-10
影响因子：
14.9
通讯作者：
Liu C
中科院分区：
文献类型：
--
作者：
Zhao D;Dong X;Jiang N;Zhang D;Liu C
G-四链体（G4）在分子治疗、诊断和传感等方面的应用潜力越来越大。ThT和NMM在大多数G4存在下都发荧光，但据报道，当ThT用作高通量测定来鉴定G4形成时，凝血酶结合适体（TBA）是已知的G4形成寡核苷酸的唯一例外。在这里，我们调查的ThT/NMM和TBA之间的相互作用，通过荧光光谱，圆二色性和分子对接模拟实验中存在或不存在的阳离子。结果表明，只有当ThT结合到平行的TBA四链体时，才能观察到大的ThT荧光增强，该四链体是在不存在阳离子的情况下由ThT诱导形成的。另一方面，只有在反平行TBA四链体的存在下，才能获得NMM荧光的极大促进，所述TBA四链体被K+或凝血酶诱导折叠。这两种探针对不同拓扑结构的TBA四链体的高选择性识别，可用于研究构象特异性G4与相关蛋白的相互作用，也可用于其他生物分子的无标记荧光传感。
G-quadruplexes (G4) have been found increasing potential in applications, such as molecular therapeutics, diagnostics and sensing. Both Thioflavin T (ThT) and N-Methyl mesoporphyrin IX (NMM) become fluorescent in the presence of most G4, but thrombin-binding aptamer (TBA) has been reported as the only exception of the known G4-forming oligonucleotides when ThT is used as a high-throughput assay to identify G4 formation. Here, we investigate the interactions between ThT/NMM and TBA through fluorescence spectroscopy, circular dichroism and molecular docking simulation experiments in the absence or presence of cations. The results display that a large ThT fluorescence enhancement can be observed only when ThT bind to the parallel TBA quadruplex, which is induced to form by ThT in the absence of cations. On the other hand, great promotion in NMM fluorescence can be obtained only in the presence of anti-parallel TBA quadruplex, which is induced to fold by K+ or thrombin. The highly selective recognition of TBA quadruplex with different topologies by the two probes may be useful to investigate the interactions between conformation-specific G4 and the associated proteins, and could also be applied in label-free fluorescent sensing of other biomolecules.
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影响因子：
14.9
作者：
Renaud de la Faverie A;Guédin A;Bedrat A;Yatsunyk LA;Mergny JL
通讯作者：
Mergny JL
DOI：
10.1016/j.bbapap.2010.04.001
发表时间：
2010-07
期刊：
Biochimica et biophysica acta
影响因子：
--
作者：
Biancalana M;Koide S
通讯作者：
Koide S
影响因子：
5.4
作者：
Avino, Anna;Portella, Guillem;Eritja, Ramon
通讯作者：
Eritja, Ramon
影响因子：
4.3
作者：
De Rache, Aurore;Kejnovska, Iva;Buess-Herman, Claudine
通讯作者：
Buess-Herman, Claudine
影响因子：
4.3
作者：
Hong, Yuning;Haeussler, Matthias;Tang, Ben Zhong
通讯作者：
Tang, Ben Zhong