title: Isolation and analysis of cell wall components from Streptococcus pneumoniae.
abstract: 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.
authors.affiliation: Centre for Bacterial Cell Biology, Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle upon Tyne NE2 4AX, UK.
authors.forename: Nhat Khai
authors.initials: NK
authors.lastname: Bui
authors.forename: Alice
authors.initials: A
authors.lastname: Eberhardt
authors.forename: Daniela
authors.initials: D
authors.lastname: Vollmer
authors.forename: Thomas
authors.initials: T
authors.lastname: Kern
authors.forename: Catherine
authors.initials: C
authors.lastname: Bougault
authors.forename: Alexander
authors.initials: A
authors.lastname: Tomasz
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Simorre
authors.forename: Waldemar
authors.initials: W
authors.lastname: Vollmer
mesh_terms: D002240:Carbohydrate Sequence; D002473:Cell Wall / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D010457:Peptidoglycan / Q000737:chemistry* / Q000302:isolation & purification*; D021241:Spectrometry, Mass, Electrospray Ionization; D013296:Streptococcus pneumoniae / Q000737:chemistry*; D013682:Teichoic Acids / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D010457:Peptidoglycan; D013682:Teichoic Acids