title: Alkyne-Functionalized Coumarin Compound for Analytic and Preparative 4-Thiouridine Labeling.
abstract: Bioconjugation of RNA is a dynamic field recently reinvigorated by a surge in research on post-transcriptional modification. This work focuses on the bioconjugation of 4-thiouridine, a nucleoside that occurs as a post-transcriptional modification in bacterial RNA and is used as a metabolic label and for cross-linking purposes in eukaryotic RNA. A newly designed coumarin compound named 4-bromomethyl-7-propargyloxycoumarin (PBC) is introduced, which exhibits remarkable selectivity for 4-thiouridine. Bearing a terminal alkyne group, it is conductive to secondary bioconjugation via "click chemistry", thereby offering a wide range of preparative and analytical options. We applied PBC to quantitatively monitor the metabolic incorporation of s4U as a label into RNA and for site-specific introduction of a fluorophore into bacterial tRNA at position 8, allowing the determination of its binding constant to an RNA-modification enzyme.
authors.affiliation: Institute of Pharmacy and Biochemistry, Johannes-Gutenberg University Mainz , Staudingerweg 5, D-55128 Mainz, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Schmid
authors.affiliation: Electrochemistry & Interfaces Group, Department of Chemistry, University of Warwick , Coventry, CV4 7AL United Kingdom.
authors.forename: Maria
authors.initials: M
authors.lastname: Adobes-Vidal
authors.affiliation: Institute of Pharmacy and Biochemistry, Johannes-Gutenberg University Mainz , Staudingerweg 5, D-55128 Mainz, Germany.
authors.forename: Mark
authors.identifier: 0000-0002-0154-0928
authors.initials: M
authors.lastname: Helm
mesh_terms: D000345:Affinity Labels / Q000138:chemical synthesis; D000480:Alkynes / Q000737:chemistry; D001665:Binding Sites; D057930:Click Chemistry; D003374:Coumarins / Q000737:chemistry*; D011485:Protein Binding; D012323:RNA Processing, Post-Transcriptional; D012329:RNA, Bacterial / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods*; D013891:Thiouridine / Q000737:chemistry
chemical_list: D000345:Affinity Labels; D000480:Alkynes; D003374:Coumarins; D012329:RNA, Bacterial; D013891:Thiouridine; C030123:coumarin