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Isolation and analysis of cell wall components from Streptococcus pneumoniae

Lookup NU author(s): Dr Nhat Khai Bui, Alice Eberhardt, Dr Daniela Vollmer, Thomas Kern, Professor Waldemar Vollmer


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The complex and heterogeneous cell wall of the pathogenic bacterium Streptococcus pneumoniae is composed of peptidoglycan and a covalently attached wall teichoic acid. The net-like peptidoglycan is formed by glycan chains that are crosslinked by short peptides. We have developed a method to purify the glycan chains, and we show that they are longer than approximately 25 disaccharide units. From purified peptidoglycan, we released 50 muropeptides that differ in the length of their peptides (tri-, tetra-, or pentapeptides with or without mono- or dipeptide branch), the degree of peptide crosslinking (monomer, dimer, or trimer), and the presence of modifications in the glycan chains (N-deacetylation, O-acetylation, or lack of GlcNAc or GlcNAc-MurNAc) or peptides (glutamic acid instead of glutamine). We also established a method to isolate wall teichoic acid chains and show that the most abundant chains have 6 or 7 repeating units. Finally, we obtained solid-state nuclear magnetic resonance spectra of whole insoluble cell walls. These novel tools will help to characterize mutant strains, cell wall-modifying enzymes, and protein-cell wall interactions. (C) 2011 Elsevier Inc. All rights reserved.

Publication metadata

Author(s): Bui NK, Eberhardt A, Vollmer D, Kern T, Bougault C, Tomasz A, Simorre JP, Vollmer W

Publication type: Article

Publication status: Published

Journal: Analytical Biochemistry

Year: 2012

Volume: 421

Issue: 2

Pages: 657-666

Print publication date: 01/12/2011

ISSN (print): 0003-2697

ISSN (electronic): 1096-0309

Publisher: Academic Press


DOI: 10.1016/j.ab.2011.11.026


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Funder referenceFunder name
European Commission through the EUR-INTAFAR