Ligands playing musical chairs with G-quadruplex DNA: A rapid and simple displacement assay for identifying selective G-quadruplex binders

被引:223
|
作者
Monchaud, D. [1 ]
Allain, C. [1 ]
Bertrand, H. [1 ]
Smargiasso, N. [2 ]
Rosu, F. [2 ]
Gabelica, V. [2 ]
De Cian, A. [3 ,4 ]
Mergny, J. -L. [3 ,4 ]
Teulade-Fichou, M. -R [1 ]
机构
[1] Ctr Univ Paris XI, CNRS, Inst Curie, Sect Rech,UMR176, F-91405 Orsay, France
[2] Univ Liege, Inst Chim, Lab Spectrometrie Masse, GIGA R, B-4000 Liege, Belgium
[3] Museum Natl Hist Nat, Acides Nucl Dynam Ciblage & Fonct Biol, INSERM, U565,USM 503, F-75005 Paris, France
[4] Museum Natl Hist Nat, Lab Regulat & Dynam Genome, CNRS, UMR5153,USM 503, F-75005 Paris, France
关键词
G-quadruplex DNA; thiazole orange; G-quadruplex ligands; fluorescent intercalator displacement;
D O I
10.1016/j.biochi.2008.02.019
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We report here the details of G4-FlD (G-quadruplex fluorescent intercalator displacement), a simple method aiming at evaluating quadruplex-DNA binding affinity and quadruplex-over duplex-DNA selectivity of putative ligands. This assay is based on the loss of fluorescence upon displacement of thiazole orange from quadruplex- and duplex-DNA matrices. The original protocol was tested using various quadruplex- and duplex-DNA targets, and with a wide panel of G-quadruplex ligands belonging to different families (i.e. from quinacridines to metallo-organic ligands) likely to display various binding modes. The reliability of the assay is further supported by comparisons with FRET-melting and ESI-MS assays. (C) 2008 Elsevier Masson SAS. All rights reserved.
引用
收藏
页码:1207 / 1223
页数:17
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