Chloroacetamide-Modified Nucleotide and RNA for Bioconjugations and Cross-Linking with RNA-Binding Proteins

0567559 - ÚOCHB 2024 RIV DE eng J - Journal Article
Brunderová, Mária - Krömer, Matouš - Vlková, Marta - Hocek, Michal
Chloroacetamide-Modified Nucleotide and RNA for Bioconjugations and Cross-Linking with RNA-Binding Proteins.
Angewandte Chemie - International Edition. Roč. 62, č. 7 (2023), č. článku e202213764. ISSN 1433-7851. E-ISSN 1521-3773
R&D Projects: GA ČR(CZ) GX20-00885X; GA MŠMT(CZ) EF16_019/0000729
Institutional support: RVO:61388963
Keywords : bioconjugations * cross-linking * modified RNA * proteins * RNA polymerases
OECD category: Organic chemistry
Impact factor: 16.6, year: 2022
Method of publishing: Open access
https://doi.org/10.1002/anie.202213764

Reactive RNA probes are useful for studying and identifying RNA-binding proteins. To that end, we designed and synthesized chloroacetamide-linked 7-deaza-ATP which was a good substrate for T7 RNA polymerase in in vitro transcription assay to synthesize reactive RNA probes bearing one or several reactive modifications. Modified RNA probes reacted with thiol-containing molecules as well as with cysteine- or histidine-containing peptides to form stable covalent products. They also reacted selectively with RNA-binding proteins to form cross-linked conjugates in high conversions thanks to proximity effect. Our modified nucleotide and RNA probes are promising tools for applications in RNA (bio)conjugations or RNA proteomics.
Permanent Link: https://hdl.handle.net/11104/0338809