title: A multifunctional bioconjugate module for versatile photoaffinity labeling and click chemistry of RNA.
abstract: A multifunctional reagent based on a coumarin scaffold was developed for derivatization of naive RNA. The alkylating agent N3BC [7-azido-4-(bromomethyl)coumarin], obtained by Pechmann condensation, is selective for uridine. N3BC and its RNA conjugates are pre-fluorophores which permits controlled modular and stepwise RNA derivatization. The success of RNA alkylation by N3BC can be monitored by photolysis of the azido moiety, which generates a coumarin fluorophore that can be excited with UV light of 320 nm. The azidocoumarin-modified RNA can be flexibly employed in structure-function studies. Versatile applications include direct use in photo-crosslinking studies to cognate proteins, as demonstrated with tRNA and RNA fragments from the MS2 phage and the HIV genome. Alternatively, the azide function can be used for further derivatization by click-chemistry. This allows e.g. the introduction of an additional fluorophore for excitation with visible light.
authors.affiliation: Institute of Pharmacy and Biochemistry, Johannes Gutenberg University Mainz, Staudinger Weg 5, D-55128 Mainz, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Kellner
authors.forename: Salifu
authors.initials: S
authors.lastname: Seidu-Larry
authors.forename: Jürgen
authors.initials: J
authors.lastname: Burhenne
authors.forename: Yuri
authors.initials: Y
authors.lastname: Motorin
authors.forename: Mark
authors.initials: M
authors.lastname: Helm
mesh_terms: D000477:Alkylating Agents / Q000138:chemical synthesis / Q000737:chemistry*; D001386:Azides / Q000138:chemical synthesis / Q000737:chemistry*; D002853:Chromatography, Liquid; D057930:Click Chemistry; D003374:Coumarins / Q000138:chemical synthesis / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry; D013058:Mass Spectrometry; D009705:Nucleosides / Q000737:chemistry; D019920:Photoaffinity Labels / Q000138:chemical synthesis / Q000737:chemistry*; D012313:RNA / Q000737:chemistry*; D014466:Ultraviolet Rays
chemical_list: C567957:7-azido-4-(bromomethyl)coumarin; D000477:Alkylating Agents; D001386:Azides; D003374:Coumarins; D005456:Fluorescent Dyes; D009705:Nucleosides; D019920:Photoaffinity Labels; D012313:RNA