Microfluidic-based G-quadruplex ligand displacement assay for alkaloid anticancer drug screening.
Some natural heterocyclic alkaloids containing planar group show potential to complex with specific promoter region of protooncogene for stabilizing the G-quadruplex (G4) structure which nowadays promises to be a target in anticancer drug design. However, in view of the polymorphic characteristics and structural complexity of heterocyclic alkaloids, it is desirable to develop high-throughput and low-consumption approach for anticancer drug screening. In this paper, an intensive study on alkaloid ligand/G4 DNA interaction has been conducted, demonstrating that the end-stacking interaction is the favorable binding mode between the oncogene-related Pu22 G4 DNA and the heterocyclic alkaloid ligand. Based on structural feasibility and energy minimization, a ligand displacement assay for screening alkaloid ligand in stabilizing the oncogene target G4 has been developed, which also helps to facilitate the assessment of drug specificity. Coupled with microfluidic-based DNAzyme-catalytic chemiluminescence detection, the approach showed the advantages of high sensitivity, high throughput with low sample and reagent consumptions.
Institute of Microanalytical Systems, Department of Chemistry, Zhejiang University, Hangzhou, 310058, China.
Haihui
H
Shen
Institute of Microanalytical Systems, Department of Chemistry, Zhejiang University, Hangzhou, 310058, China.
Bo
B
Zhang
Institute of Microanalytical Systems, Department of Chemistry, Zhejiang University, Hangzhou, 310058, China.
Huiyan
H
Xu
Institute of Microanalytical Systems, Department of Chemistry, Zhejiang University, Hangzhou, 310058, China.
Yue
Y
Sun
Institute of Microanalytical Systems, Department of Chemistry, Zhejiang University, Hangzhou, 310058, China.
Qiwang
Q
Wu
Institute of Microanalytical Systems, Department of Chemistry, Zhejiang University, Hangzhou, 310058, China. Electronic address: shzju@zju.edu.cn.
Hong
H
Shen
Institute of Microanalytical Systems, Department of Chemistry, Zhejiang University, Hangzhou, 310058, China. Electronic address: liuyingch@zju.edu.cn.
Yingchun
Y
Liu
D000470:Alkaloids / Q000032:analysis*; D000970:Antineoplastic Agents / Q000032:analysis*; D021881:DNA, Catalytic / Q000737:chemistry; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004354:Drug Screening Assays, Antitumor / Q000379:methods; D054856:G-Quadruplexes; D008024:Ligands; D008163:Luminescent Measurements / Q000379:methods*; D044085:Microfluidics / Q000379:methods*
D000470:Alkaloids; D000970:Antineoplastic Agents; D021881:DNA, Catalytic; D008024:Ligands
Alkaloid; Chemiluminescence; Drug screening; G-quadruplex; Ligand displacement assay