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 di Guilmi, A. M.
Right arrow Articles by Vernet, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by di Guilmi, A. M.
Right arrow Articles by Vernet, T.

 Previous Article  |  Next Article 

Journal of Bacteriology, May 1999, p. 2773-2781, Vol. 181, No. 9
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Glycosyltransferase Domain of Penicillin-Binding Protein 2a from Streptococcus pneumoniae Is Membrane Associateddagger

Anne Marie di Guilmi,1 Nicolas Mouz,1 Lydie Martin,1 JoAnn Hoskins,2 S. Richard Jaskunas,2 Otto Dideberg,1 and Thierry Vernet1,*

Institut de Biologie Structurale Jean-Pierre Ebel (CEA/CNRS), 38027 Grenoble Cedex 1, France,1 and Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana 46285-043822

Received 23 December 1998/Accepted 26 February 1999

Penicillin-binding proteins (PBPs) are bacterial cytoplasmic membrane proteins that catalyze the final steps of the peptidoglycan synthesis. Resistance to beta -lactams in Streptococcus pneumoniae is caused by low-affinity PBPs. S. pneumoniae PBP 2a belongs to the class A high-molecular-mass PBPs having both glycosyltransferase (GT) and transpeptide (TP) activities. Structural and functional studies of both domains are required to unravel the mechanisms of resistance, a prerequisite for the development of novel antibiotics. The extracellular region of S. pneumoniae PBP 2a has been expressed (PBP 2a*) in Escherichia coli as a glutathione S-transferase fusion protein. The acylation kinetic parameters of PBP 2a* for beta -lactams were determined by stopped-flow fluorometry. The acylation efficiency toward benzylpenicillin was much lower than that toward cefotaxime, a result suggesting that PBP 2a participates in resistance to cefotaxime and other beta -lactams, but not in resistance to benzylpenicillin. The TP domain was purified following limited proteolysis. PBP 2a* required detergents for solubility and interacted with lipid vesicles, while the TP domain was water soluble. We propose that PBP 2a* interacts with the cytoplasmic membrane in a region distinct from its transmembrane anchor region, which is located between Lys 78 and Ser 156 of the GT domain.

* Corresponding author. Mailing address: Institut de Biologie Structurale Jean-Pierre Ebel, Laboratoire d'Ingénierie des Macromolécules, 41 Avenue des Martyrs, 38027 Grenoble Cedex 1, France. Phone: 33-04 76 88 96 81. Fax: 33-04 76 88 54 94. E-mail: vernet{at}ibs.fr.

dagger This is publication no. 630 of the Institut de Biologie Structurale Jean-Pierre Ebel.

Journal of Bacteriology, May 1999, p. 2773-2781, Vol. 181, No. 9
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • van Heijenoort, J. (2007). Lipid Intermediates in the Biosynthesis of Bacterial Peptidoglycan. Microbiol. Mol. Biol. Rev. 71: 620-635 [Abstract] [Full Text]  
  • Haenni, M., Moreillon, P. (2006). Mutations in Penicillin-Binding Protein (PBP) Genes and in Non-PBP Genes during Selection of Penicillin-Resistant Streptococcus gordonii. Antimicrob. Agents Chemother. 50: 4053-4061 [Abstract] [Full Text]  
  • Haenni, M., Majcherczyk, P. A., Barblan, J.-L., Moreillon, P. (2006). Mutational Analysis of Class A and Class B Penicillin-Binding Proteins in Streptococcus gordonii. Antimicrob. Agents Chemother. 50: 4062-4069 [Abstract] [Full Text]  
  • Terrak, M., Nguyen-Disteche, M. (2006). Kinetic Characterization of the Monofunctional Glycosyltransferase from Staphylococcus aureus.. J. Bacteriol. 188: 2528-2532 [Abstract] [Full Text]  
  • Morlot, C., Pernot, L., Le Gouellec, A., Di Guilmi, A. M., Vernet, T., Dideberg, O., Dessen, A. (2005). Crystal Structure of a Peptidoglycan Synthesis Regulatory Factor (PBP3) from Streptococcus pneumoniae. J. Biol. Chem. 280: 15984-15991 [Abstract] [Full Text]  
  • Duez, C., Hallut, S., Rhazi, N., Hubert, S., Amoroso, A., Bouillenne, F., Piette, A., Coyette, J. (2004). The ponA Gene of Enterococcus faecalis JH2-2 Codes for a Low-Affinity Class A Penicillin-Binding Protein. J. Bacteriol. 186: 4412-4416 [Abstract] [Full Text]  
  • Pagliero, E., Chesnel, L., Hopkins, J., Croize, J., Dideberg, O., Vernet, T., Di Guilmi, A. M. (2004). Biochemical Characterization of Streptococcus pneumoniae Penicillin-Binding Protein 2b and Its Implication in {beta}-Lactam Resistance. Antimicrob. Agents Chemother. 48: 1848-1855 [Abstract] [Full Text]  
  • Di Guilmi, A. M., Dessen, A., Dideberg, O., Vernet, T. (2003). The Glycosyltransferase Domain of Penicillin-Binding Protein 2a from Streptococcus pneumoniae Catalyzes the Polymerization of Murein Glycan Chains. J. Bacteriol. 185: 4418-4423 [Abstract] [Full Text]  
  • Di Guilmi, A. M., Dessen, A., Dideberg, O., Vernet, T. (2003). Functional Characterization of Penicillin-Binding Protein 1b from Streptococcus pneumoniae. J. Bacteriol. 185: 1650-1658 [Abstract] [Full Text]  
  • Heijenoort, J. v. (2001). Formation of the glycan chains in the synthesis of bacterial peptidoglycan. Glycobiology 11: 25R-36R [Abstract] [Full Text]  
  • Zhao, G., Meier, T. I., Hoskins, J., McAllister, K. A. (2000). Identification and Characterization of the Penicillin-Binding Protein 2a of Streptococcus pneumoniae and Its Possible Role in Resistance to beta -Lactam Antibiotics. Antimicrob. Agents Chemother. 44: 1745-1748 [Abstract] [Full Text]  
  • Dessen, A., Mouz, N., Gordon, E., Hopkins, J., Dideberg, O. (2001). Crystal Structure of PBP2x from a Highly Penicillin-resistant Streptococcus pneumoniae Clinical Isolate. A MOSAIC FRAMEWORK CONTAINING 83 MUTATIONS. J. Biol. Chem. 276: 45106-45112 [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»