Labelling of nucleosides and oligonucleotides by solvatochromic 4-aminophthalimide fluorophore for studying DNA–protein interactions

0382144 - ÚOCHB 2013 RIV GB eng J - Journal Article
Riedl, Jan - Pohl, Radek - Ernsting, N. P. - Orság, Petr - Fojta, Miroslav - Hocek, Michal
Labelling of nucleosides and oligonucleotides by solvatochromic 4-aminophthalimide fluorophore for studying DNA–protein interactions.
Chemical Science. Roč. 3, č. 9 (2012), s. 2797-2806. ISSN 2041-6520. E-ISSN 2041-6539
R&D Projects: GA ČR GA203/09/0317; GA ČR GBP206/12/G151
Institutional support: RVO:61388963 ; RVO:68081707
Keywords : DNA * oligonucleotides * polymerase * phthalimide * nucleotides * fluorescent labeling
Subject RIV: CC - Organic Chemistry
Impact factor: 8.314, year: 2012

Solvatochromic fluorescent 4-aminophthalimide and 4-(N,N-dimethylamino)phthalimide were attached covalently to 2'-deoxycytidine or -adenosine via non-conjugated propargyl linker by Sonogashira cross-coupling reactions. The nucleosides were phosphorylated to triphosphates and enzymatically incorporated to oligonucleotides by DNA polymerases. API-labelled DNA was used for detection of DNA protein interactions with p53 protein or single strand binding protein. Both proteins changed the polarity around the fluorophore and increased the intensity of API fluorescence.
Permanent Link: http://hdl.handle.net/11104/0212451