Publication details

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

Authors

RIEDL J POHL R ERNSTING NP ORSÁG Petr FOJTA Miroslav HOCEK Michal

Year of publication 2012
Type Article in Periodical
Magazine / Source CHEMICAL SCIENCE
Citation
Doi http://dx.doi.org/10.1039/c2sc20404e
Description Solvatochromic fluorescent 4-aminophthalimide (API) and 4-(N, N-dimethylamino) phthalimide (DAPI) were attached covalently to 2'-deoxycytidine or -adenosine via a non-conjugated propargyl linker by Sonogashira cross-coupling reactions of N-propargylphthalimides with halogenated nucleosides. The nucleosides were phosphorylated to triphosphates and enzymatically incorporated into oligonucleotides by DNA polymerases. API-labelled DNA was used for the detection of DNA protein interactions with either the sequence-specific p53 protein or a non-specific single strand binding (SSB) protein. Both proteins changed the polarity around the fluorophore and increased (2-3 fold) the intensity of API fluorescence.

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