title: Efficient access to peptidyl-RNA conjugates for picomolar inhibition of non-ribosomal FemX(Wv) aminoacyl transferase.
abstract: Peptidyl-RNA conjugates have various applications in studying the ribosome and enzymes participating in tRNA-dependent pathways such as Fem transferases in peptidoglycan synthesis. Herein a convergent synthesis of peptidyl-RNAs based on Huisgen-Sharpless cycloaddition for the final ligation step is developed. Azides and alkynes are introduced into tRNA and UDP-MurNAc-pentapeptide, respectively. Synthesis of 2'-azido RNA helix starts from 2'-azido-2'-deoxyadenosine that is coupled to deoxycytidine by phosphoramidite chemistry. The resulting dinucleotide is deprotected and ligated to a 22-nt RNA helix mimicking the acceptor arm of Ala-tRNA(Ala) by T4 RNA ligase. For alkyne UDP-MurNAc-pentapeptide, meso-cystine is enzymatically incorporated into the peptidoglycan precursor and reduced, and L-Cys is converted to dehydroalanine with O-(mesitylenesulfonyl)hydroxylamine. Reaction of but-3-yne-1-thiol with dehydroalanine affords the alkyne-containing UDP-MurNAc-pentapeptide. The Cu(I)-catalyzed azide alkyne cycloaddition reaction in the presence of tris[(1-hydroxypropyl-1H-1,2,3-triazol-4-yl)methyl]amine provided the peptidyl-RNA conjugate, which was tested as an inhibitor of non-ribosomal FemX(Wv) aminoacyl transferase. The bi-substrate analogue was found to inhibit FemX(Wv) with an IC(50) of (89±9) pM, as both moieties of the peptidyl-RNA conjugate contribute to high-affinity binding.
authors.affiliation: Centre de Recherche des Cordeliers, LRMA, Equipe 12, Université Pierre et Marie Curie - Paris 6, UMR S 872, Paris 75006, France.
authors.forename: Matthieu
authors.initials: M
authors.lastname: Fonvielle
authors.forename: Dénia
authors.initials: D
authors.lastname: Mellal
authors.forename: Delphine
authors.initials: D
authors.lastname: Patin
authors.forename: Maxime
authors.initials: M
authors.lastname: Lecerf
authors.forename: Didier
authors.initials: D
authors.lastname: Blanot
authors.forename: Ahmed
authors.initials: A
authors.lastname: Bouhss
authors.forename: Marco
authors.initials: M
authors.lastname: Santarem
authors.forename: Dominique
authors.initials: D
authors.lastname: Mengin-Lecreulx
authors.forename: Matthieu
authors.initials: M
authors.lastname: Sollogoub
authors.forename: Michel
authors.initials: M
authors.lastname: Arthur
authors.forename: Mélanie
authors.initials: M
authors.lastname: Ethève-Quelquejeu
mesh_terms: D019881:Aminoacyltransferases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002384:Catalysis; D003300:Copper / Q000737:chemistry; D061565:Cycloaddition Reaction; D007700:Kinetics; D009842:Oligopeptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D012313:RNA / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D011130:RNA Ligase (ATP) / Q000378:metabolism; D014538:Uridine Diphosphate N-Acetylmuramic Acid / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism
chemical_list: D009842:Oligopeptides; D014538:Uridine Diphosphate N-Acetylmuramic Acid; C002225:UDP-N-acetylmuramic acid pentapeptide; D012313:RNA; D003300:Copper; D019881:Aminoacyltransferases; D011130:RNA Ligase (ATP)