This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Maillard, A. P.
Right arrow Articles by Arthur, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Maillard, A. P.
Right arrow Articles by Arthur, M.

 Previous Article  |  Next Article 

Journal of Bacteriology, June 2005, p. 3833-3838, Vol. 187, No. 11
0021-9193/05/$08.00+0     doi:10.1128/JB.187.11.3833-3838.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Structure-Based Site-Directed Mutagenesis of the UDP-MurNAc-Pentapeptide-Binding Cavity of the FemX Alanyl Transferase from Weissella viridescens

Antoine P. Maillard ,1,{dagger},{ddagger} Sabrina Biarrotte-Sorin,1,2,{ddagger} Régis Villet,1 Stéphane Mesnage,1 Ahmed Bouhss,3 Wladimir Sougakoff,1 Claudine Mayer,1 and Michel Arthur1*

Laboratoire de Recherche Moléculaire sur les Antibiotiques, INSERM U655, Université Paris 6,1 Laboratoire de Minéralogie-Cristallographie de Paris, Université Paris 6, Paris,2 Enveloppes Bactériennes et Antibiotiques CNRS, UMR 8619, Université Paris 11, Orsay, France3

Received 20 December 2004/ Accepted 23 February 2005

Weissella viridescens FemX (FemXWv) belongs to the Fem family of nonribosomal peptidyl transferases that use aminoacyl-tRNA as the amino acid donor to synthesize the peptide cross-bridge found in the peptidoglycan of many species of pathogenic gram-positive bacteria. We have recently solved the crystal structure of FemXWv in complex with the peptidoglycan precursor UDP-MurNAc-pentapeptide and report here the site-directed mutagenesis of nine residues located in the binding cavity for this substrate. Two substitutions, Lys36Met and Arg211Met, depressed FemXWv transferase activity below detectable levels without affecting protein folding. Analogues of UDP-MurNAc-pentapeptide lacking the phosphate groups or the C-terminal D-alanyl residues were not substrates of the enzyme. These results indicate that Lys36 and Arg211 participate in a complex hydrogen bond network that connects the C-terminal D-Ala residues to the phosphate groups of UDP-MurNAc-pentapeptide and constrains the substrate in a conformation that is essential for transferase activity.

* Corresponding author. Mailing address: LRMA INSERM U655, Université Paris 6, 15 rue de l'Ecole de Médecine, 75270 Paris Cedex 06, France. Phone: 33 1 43 25 00 33. Fax: 33 1 43 25 68 12. E-mail: michel.arthur{at}bhdc.jussieu.fr.

{dagger} Present address: Faculty of Medicine, Department of Biochemistry, University of British Columbia, V6T 1Z3 Vancouver, Canada.

{ddagger} A.P.M. and S.B.-S. contributed equally to this work.

Journal of Bacteriology, June 2005, p. 3833-3838, Vol. 187, No. 11
0021-9193/05/$08.00+0     doi:10.1128/JB.187.11.3833-3838.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Fonvielle, M., Chemama, M., Villet, R., Lecerf, M., Bouhss, A., Valery, J.-M., Etheve-Quelquejeu, M., Arthur, M. (2009). Aminoacyl-tRNA recognition by the FemXWv transferase for bacterial cell wall synthesis. Nucleic Acids Res 37: 1589-1601 [Abstract] [Full Text]  
  • De Pascale, G., Lloyd, A. J., Schouten, J. A., Gilbey, A. M., Roper, D. I., Dowson, C. G., Bugg, T. D. H. (2008). Kinetic Characterization of Lipid II-Ala:Alanyl-tRNA Ligase (MurN) from Streptococcus pneumoniae using Semisynthetic Aminoacyl-lipid II Substrates. J. Biol. Chem. 283: 34571-34579 [Abstract] [Full Text]  
  • Garg, R. P., Qian, X. L., Alemany, L. B., Moran, S., Parry, R. J. (2008). Investigations of valanimycin biosynthesis: Elucidation of the role of seryl-tRNA. Proc. Natl. Acad. Sci. USA 105: 6543-6547 [Abstract] [Full Text]  
  • Lloyd, A. J., Gilbey, A. M., Blewett, A. M., De Pascale, G., El Zoeiby, A., Levesque, R. C., Catherwood, A. C., Tomasz, A., Bugg, T. D. H., Roper, D. I., Dowson, C. G. (2008). Characterization of tRNA-dependent Peptide Bond Formation by MurM in the Synthesis of Streptococcus pneumoniae Peptidoglycan. J. Biol. Chem. 283: 6402-6417 [Abstract] [Full Text]  
  • van Heijenoort, J. (2007). Lipid Intermediates in the Biosynthesis of Bacterial Peptidoglycan. Microbiol. Mol. Biol. Rev. 71: 620-635 [Abstract] [Full Text]  
  • Villet, R., Fonvielle, M., Busca, P., Chemama, M., Maillard, A. P., Hugonnet, J.-E., Dubost, L., Marie, A., Josseaume, N., Mesnage, S., Mayer, C., Valery, J.-M., Etheve-Quelquejeu, M., Arthur, M. (2007). Idiosyncratic features in tRNAs participating in bacterial cell wall synthesis. Nucleic Acids Res 35: 6870-6883 [Abstract] [Full Text]  
  • El Ghachi, M., Bouhss, A., Barreteau, H., Touze, T., Auger, G., Blanot, D., Mengin-Lecreulx, D. (2006). Colicin M Exerts Its Bacteriolytic Effect via Enzymatic Degradation of Undecaprenyl Phosphate-linked Peptidoglycan Precursors. J. Biol. Chem. 281: 22761-22772 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»