Assessment of selectivity of G-quadruplex ligands via an optimised FRET melting assay
Assessment of selectivity of G-quadruplex ligands via an optimised FRET melting assay
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DOI：
10.1016/j.biochi.2015.06.002
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发表时间：
2015-08-01
期刊：
影响因子：
3.9
通讯作者：
Mergny, Jean-Louis
中科院分区：
文献类型：
--
作者：
De Rache, Aurore;Mergny, Jean-Louis
四链G-四链体（G4）DNA结构是有前途的药物靶点，因为这些非经典结构似乎可以调节基因表达和端粒生长。虽然所有类型的G4都是由四个鸟苷四联体稳定的，但环、沟和侧翼碱基的差异导致G4结构之间令人印象深刻的结构多样性，这可能允许小分子配体的选择性识别。我们适应了先前描述的福斯特共振能量转移（FRET）熔解试验，以评估不同的G-四链体拓扑结构的G4配体的选择性。我们证明了FAM和塔姆拉荧光染料的掺入和PEG的存在影响了具有G-四链体形成潜力的某些序列所采用的结构。测量条件的优化确保了所选的一组G4 DNA寡核苷酸在有和没有配体的可测量温度范围内的折叠和热稳定性。优化的方法能够比较众所周知的G4配体，如TmPyP 4，Braco 19，pyridostatin，360 A，PhenDC 3和TrisQ。(C)2015 Elsevier B. V.和法国生物化学与生物分子学会（SFBBM）。All rights reserved.
Four-stranded G-quadruplex (G4) DNA structures are promising drug targets as these non-canonical structures appear to regulate gene expression and telomere growth. Although all types of G4 are stabilised by quartets of four guanosines, differences in loops, grooves, and flanking bases, result in an impressive structural diversity among G4 structures that may allow selective recognition by small molecule ligands. We adapted the previously described Forster resonance energy transfer (FRET) melting assay to evaluate the selectivity of G4 ligands for different G-quadruplex topologies. We demonstrated that the incorporation of FAM and Tamra fluorescent dyes and the presence of PEG influenced the structures adopted by certain sequences with G-quadruplex-forming potential. Optimisation of the measurement conditions ensured the folding and thermal stability of a selected set of G4 DNA oligonucleotides in a measurable temperature range with and without ligand. The optimised method enabled comparison of well known G4 ligands such as TmPyP4, Braco19, pyridostatin, 360A, PhenDC3, and TrisQ. (C) 2015 Elsevier B.V. and Societe Francaise de Biochimie et Biologie Moleculaire (SFBBM). All rights reserved.