Unit

UNIT 4.44 Synthesis of Stable Aminoacyl-tRNA Analogs

  1. Maryline Chemama1,
  2. Matthieu Fonvielle2,
  3. Maxime Lecerf2,
  4. Dénia Mellal1,
  5. Hélène Fief1,
  6. Michel Arthur2,
  7. Mélanie Etheve-Quelquejeu1

Published Online: 1 MAR 2011

DOI: 10.1002/0471142700.nc0444s44

Current Protocols in Nucleic Acid Chemistry

Current Protocols in Nucleic Acid Chemistry

How to Cite

Chemama, M., Fonvielle, M., Lecerf, M., Mellal, D., Fief, H., Arthur, M. and Etheve-Quelquejeu, M. 2011. Synthesis of Stable Aminoacyl-tRNA Analogs. Current Protocols in Nucleic Acid Chemistry. 44:4.44:4.44.1–4.44.33.

Author Information

  1. 1

    Institut Parisien de Chimie Moléculaire, Université Pierre et Marie Curie, Paris, France

  2. 2

    Laboratoire de Recherche Moléculaire sur les Antibiotiques, Centre de Recherche des Cordeliers, Université Pierre et Marie Curie, Paris, France

Publication History

  1. Published Online: 1 MAR 2011
  2. Published Print: MAR 2011

Abstract

Aminoacyl-tRNAs have important roles in a variety of biological processes. Here, we describe the synthesis of stable aminoacyl-tRNA analogs containing 1,4-substituted 1,2,3-triazole rings. The procedure involves (i) copper-catalyzed cycloadditions of 3′-or 2′-azido-adenosine and alkynes, (ii) coupling between the resulting triazole-deoxyadenosine derivatives and a deoxycytidine phosphoramidite, and (iii) the enzymatic ligation of the 2′- or 3′-triazole-dinucleotides with a 22-nt RNA microhelix that mimics the acceptor arm of tRNA. Each nucleoside and nucleotide intermediate was characterized by MS spectrometry and 1H, 31P, and 13C NMR spectroscopy, and the tRNA-analogs were assayed for inhibition of FemXWv, an alanyl-transferase essential for the formation of the peptidoglycan network of Gram-positive bacterial pathogens. The low IC50 values obtained (2 to 4 µM) indicate that the five-membered triazole rings acted as an isosteres of esters and can be used for the design of stable aminoacyl-tRNA analogs. Curr. Protoc. Nucleic Acid Chem. 44:4.44.1-4.44.33. © 2011 by John Wiley & Sons, Inc.

Keywords:

  • aminoacyl-tRNA;
  • click chemistry;
  • dinucleotides;
  • inhibitors;
  • triazoles;
  • azidoadenosine;
  • transferase