李卉卉副教授、David Chen教授和楊小弟教授課題組合作在TALANTA發(fā)表研究論文
Evaluation of the binding of natural products with thrombin binding aptamer G-quadruplex using electrospray ionization mass spectrometry and spectroscopic methods
Yang, PF (Yang, Pengfei)[ 1 ] ; Wang, XY (Wang, Xinyi)[ 1 ] ; Gu, ZG (Gu, Zhenggui)[ 1 ] ; Li, H (Li, Huihui)[ 1 ]*（李卉卉）; Chen, DDY (Chen, David D. Y.)[ 2 ]*（David Chen） ; Yang, XD (Yang, Xiaodi)[ 1 ]*（楊小弟）
[ 1 ] Nanjing Normal Univ, Sch Chem & Mat Sci, Jiangsu Key Lab Biomed Mat, Jiangsu Collaborat Innovat Ctr Biomed Funct Mat, Nanjing 210023, Jiangsu, Peoples R China
[ 2 ] Univ British Columbia, Dept Chem, Vancouver, BC V6T 1Z1, Canada
TALANTA，201908,200,424-431
A 15-mer thrombin-binding aptamer (TBA) was discovered with specificity for thrombin. It forms a unique G-quadruplex (G4), which is postulated to be the molecular basis for its binding specificity. Many analytical methods make use of affinity binding between the thrombin and TBA as they form a very stable complex. We develop a strategy to stabilize TBA/G4's structure by introducing G4-interactive molecules, which may enhance its ability to recognize the target. Herein, a fast screening ESI-MS assay was employed to determine potential binding of natural products molecules with the TBA/G4 complex. The experimental results showed that four investigated natural alkaloids had apparent binding affinities. One of them, jatrorrhizine (L1), has been shown to bind strongly to the TBA/G4 mainly in 1:2 M ratio. Once the working conditions were established, the interaction of the jatrorrhizine with the TBA/G4 was explored using a combination of ESI-MS and spectroscopic techniques. Ligand-induced effects on TBA/G4 structure and its stability were examined by means of circular dichroism (CD). Jatrorrhizine inducing the G4 formation seems also to be the more effective in terms of thermal stabilization under the experimental conditions used. Both results of UV and fluorescence experiments undoubtedly showed a good binding affinity with the binding constant around 10(5) L mol(-1). The stacking interactions of jatrorrhizine with the G-tetrads in TBA/G4 were further confirmed by competition experiment. ESI-MS was carried out to determine the coexistence of 1:1 and 1:2 complexes in TBA/G4-L1 system, and showed a dynamical shift from 1:1 to 1:2 complex in minutes.
文章鏈接：
https://www.sciencedirect.com/science/article/pii/S0039914019303376?via%3Dihub
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