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

RIEDL, J, R POHL, NP ERNSTING, Petr ORSÁG, Miroslav FOJTA and Michal HOCEK. Labelling of nucleosides and oligonucleotides by solvatochromic 4-aminophthalimide fluorophore for studying DNA-protein interactions. CHEMICAL SCIENCE. Cambridge: ROYAL SOC CHEMISTRY, 2012, vol. 3, No 9, p. 2797-2806. ISSN 2041-6520. Available from: https://dx.doi.org/10.1039/c2sc20404e.

Basic information

• Original name: Labelling of nucleosides and oligonucleotides by solvatochromic 4-aminophthalimide fluorophore for studying DNA-protein interactions
• Authors: RIEDL, J, R POHL, NP ERNSTING, Petr ORSÁG, Miroslav FOJTA and Michal HOCEK.
• Edition: CHEMICAL SCIENCE, Cambridge, ROYAL SOC CHEMISTRY, 2012, 2041-6520.

Other information

• Original language: English
• Type of outcome: Article in a journal
• Confidentiality degree: is not subject to a state or trade secret
• Impact factor: Impact factor: 8.314
• Doi: http://dx.doi.org/10.1039/c2sc20404e
• UT WoS: 000306931300019

Abstract

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.