title: Native Chemical Ligation of Hydrolysis-Resistant 3'-NH-Cysteine-Modified RNA.
abstract: Hydrolysis-resistant RNA-peptide conjugates that contain a 3'-NH linkage between the adenosine ribose and the C-terminal carboxyl group of a peptide moiety instead of the natural ester mimic acylated tRNA termini. Their detailed preparation that combines solid-phase oligonucleotide synthesis and bioconjugation is described here. The key step is native chemical ligation (NCL) of 3'-NH-cysteine-modified RNA to highly soluble peptide thioesters. These hydrolysis-resistant 3'-NH-peptide-modified RNAs, containing the universally conserved 3'-CCA end of tRNA, are biologically active and can bind to the ribosome. They can be used as valuable probes for structural and functional studies of the ribosomal elongation cycle.
authors.affiliation: Department of Chemistry, Graduate School of Science, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.
authors.forename: Anna-Skrollan
authors.initials: AS
authors.lastname: Geiermann
authors.affiliation: Institute of Organic Chemistry, Center for Chemistry and Biomedicine, Leopold Franzens University, Innsbruck, Austria.
authors.forename: Ronald
authors.initials: R
authors.lastname: Micura
mesh_terms: D003545:Cysteine / Q000737:chemistry; D006868:Hydrolysis; D009841:Oligonucleotides / Q000737:chemistry; D010455:Peptides / Q000737:chemistry*
chemical_list: D009841:Oligonucleotides; D010455:Peptides; D003545:Cysteine
keywords: RNA solid-phase synthesis; native chemical ligation; nucleoside modification; oligonucleotide-peptide conjugates; peptides