Efficient Access to Peptidyl-RNA Conjugates for Picomolar Inhibition of Non-ribosomal FemXWv Aminoacyl Transferase

Authors

  • Dr. Matthieu Fonvielle ,

    1. Centre de Recherche des Cordeliers, LRMA, Equipe 12, Université Pierre et Marie Curie - Paris 6, UMR S 872, Paris 75006 (France), Université Paris Descartes, Sorbonne Paris Cité, UMR S 872, Paris 75006 (France), INSERM, U872, Paris, 75006 (France), Fax: (+33) 1-4325-6812
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    • These authors contributed equally to this work.

  • Dénia Mellal ,

    1. Institut Parisien de Chimie Moléculaire, CNRS UMR 7201, Université Pierre et Marie Curie - Paris 6, 4, place Jussieu, 75005 Paris (France), Fax: (+33) 1-4427-5504
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    • These authors contributed equally to this work.

  • Delphine Patin,

    1. Laboratoire des Enveloppes Bactériennes et Antibiotiques, Institut de Biochimie et de Biophysique Moléculaire et Cellulaire, UMR 8619, CNRS, Université Paris-Sud, 91405 Orsay (France)
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  • Maxime Lecerf,

    1. Centre de Recherche des Cordeliers, LRMA, Equipe 12, Université Pierre et Marie Curie - Paris 6, UMR S 872, Paris 75006 (France), Université Paris Descartes, Sorbonne Paris Cité, UMR S 872, Paris 75006 (France), INSERM, U872, Paris, 75006 (France), Fax: (+33) 1-4325-6812
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  • Dr. Didier Blanot,

    1. Laboratoire des Enveloppes Bactériennes et Antibiotiques, Institut de Biochimie et de Biophysique Moléculaire et Cellulaire, UMR 8619, CNRS, Université Paris-Sud, 91405 Orsay (France)
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  • Dr. Ahmed Bouhss,

    1. Laboratoire des Enveloppes Bactériennes et Antibiotiques, Institut de Biochimie et de Biophysique Moléculaire et Cellulaire, UMR 8619, CNRS, Université Paris-Sud, 91405 Orsay (France)
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  • Dr. Marco Santarem,

    1. Institut Parisien de Chimie Moléculaire, CNRS UMR 7201, Université Pierre et Marie Curie - Paris 6, 4, place Jussieu, 75005 Paris (France), Fax: (+33) 1-4427-5504
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  • Dr. Dominique Mengin-Lecreulx,

    1. Laboratoire des Enveloppes Bactériennes et Antibiotiques, Institut de Biochimie et de Biophysique Moléculaire et Cellulaire, UMR 8619, CNRS, Université Paris-Sud, 91405 Orsay (France)
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  • Dr. Matthieu Sollogoub,

    1. Institut Parisien de Chimie Moléculaire, CNRS UMR 7201, Université Pierre et Marie Curie - Paris 6, 4, place Jussieu, 75005 Paris (France), Fax: (+33) 1-4427-5504
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  • Dr. Michel Arthur,

    Corresponding author
    1. Centre de Recherche des Cordeliers, LRMA, Equipe 12, Université Pierre et Marie Curie - Paris 6, UMR S 872, Paris 75006 (France), Université Paris Descartes, Sorbonne Paris Cité, UMR S 872, Paris 75006 (France), INSERM, U872, Paris, 75006 (France), Fax: (+33) 1-4325-6812
    • Centre de Recherche des Cordeliers, LRMA, Equipe 12, Université Pierre et Marie Curie - Paris 6, UMR S 872, Paris 75006 (France), Université Paris Descartes, Sorbonne Paris Cité, UMR S 872, Paris 75006 (France), INSERM, U872, Paris, 75006 (France), Fax: (+33) 1-4325-6812
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  • Dr. Mélanie Ethève-Quelquejeu

    Corresponding author
    1. Institut Parisien de Chimie Moléculaire, CNRS UMR 7201, Université Pierre et Marie Curie - Paris 6, 4, place Jussieu, 75005 Paris (France), Fax: (+33) 1-4427-5504
    • Institut Parisien de Chimie Moléculaire, CNRS UMR 7201, Université Pierre et Marie Curie - Paris 6, 4, place Jussieu, 75005 Paris (France), Fax: (+33) 1-4427-5504
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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-tRNAAla 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 CuI-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 FemXWv aminoacyl transferase. The bi-substrate analogue was found to inhibit FemXWv with an IC50 of (89±9) pM, as both moieties of the peptidyl–RNA conjugate contribute to high-affinity binding.

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