Alkyne-Functionalized Coumarin Compound for Analytic and Preparative 4-Thiouridine Labeling

作者：Schmid Katharina; Adobes Vidal Maria; Helm Mark

来源：Bioconjugate Chemistry, 2017, 28(4): 1123-1134.

DOI：10.1021/acs.bioconjchem.7b00035

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-bromomethy1-7-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 s(4)U 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.

• 出版日期2017-4