0603499 - ÚOCHB 2026 RIV US eng J - Journal Article
Ivancová, Ivana - Sánchez Quirante, Tania - Ondruš, Marek - Pohl, Radek - Vlková, Marta - Žilecká, Eva - Bouřa, Evžen - Hocek, Michal
Enzymatic synthesis of reactive RNA probes containing squaramate-linked cytidine or adenosine for bioconjugations and cross-linking with lysine-containing peptides and proteins.
Communications Chemistry. Roč. 8, January (2025), č. článku 1. ISSN 2399-3669. E-ISSN 2399-3669
R&D Projects: GA ČR(CZ) GX20-00885X; GA MŠMT(CZ) EH22_008/0004575
Institutional support: RVO:61388963
Keywords : binding sites * DNA * identification
OECD category: Organic chemistry
Impact factor: 5.9, year: 2023 ; AIS: 1.585, rok: 2023
Method of publishing: Open access
Result website:
https://doi.org/10.1038/s42004-024-01399-6DOI: https://doi.org/10.1038/s42004-024-01399-6

Protein-RNA interactions play important biological roles and hence reactive RNA probes for cross-linking with proteins are important tools in their identification and study. To this end, we designed and synthesized 5′-O-triphosphates bearing a reactive squaramate group attached to position 5 of cytidine or position 7 of 7-deazaadenosine and used them as substrates for polymerase synthesis of modified RNA. In vitro transcription with T7 RNA polymerase or primer extension using TGK polymerase was used for synthesis of squaramate-modified RNA probes which underwent covalent bioconjugations with amine-linked fluorophore and lysine-containing peptides and proteins including several viral RNA polymerases or HIV reverse transcriptase. Inhibition of RNA-depending RNA polymerases from Japanese Encephalitis virus was observed through formation of covalent cross-link which was partially identified by MS/MS analysis. Thus, the squaramate-linked NTP analogs are useful building blocks for the synthesis of reactive RNA probes for bioconjugations with primary amines and cross-linking with lysine residues.
Permanent Link: https://hdl.handle.net/11104/0360822
Research data: Národní repozitář