Analysis of cell wall teichoic acids in Staphylococcus aureus

Gonçalo Covas, Filipa Vaz, Gabriela Henriques, Mariana G. Pinho, Sérgio R. Filipe

Research output: Chapter in Book/Report/Conference proceedingChapter

5 Citations (Scopus)

Abstract

Most bacterial cells are surrounded by a surface composed mainly of peptidoglycan (PGN), a glycopolymer responsible for ensuring the bacterial shape and a telltale molecule that betrays the presence of bacteria to the host immune system. In Staphylococcus aureus, as in most gram-positive bacteria, peptidoglycan is concealed by covalently linked molecules of wall teichoic acids (WTA)—phosphate rich molecules made of glycerol and ribitol phosphates which may be tailored by different amino acids and sugars. In order to analyze and compare the composition of WTA produced by different S. aureus strains, we describe methods to: (1) quantify the total amount of WTA present at the bacterial cell surface, through the determination of the inorganic phosphate present in phosphodiester linkages of WTA; (2) identify which sugar constituents are present in the assembled WTA molecules, by detecting the monosaccharides, released by acid hydrolysis, through an high-performance anion exchange chromatography analysis coupled with pulsed amperometric detection (HPAEC-PAD) and (3) compare the polymerization degree of WTA found at the cell surface of different S. aureus strains, through their different migration in a polyacrylamide gel electrophoresis (PAGE).

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages201-213
Number of pages13
Volume1440
DOIs
Publication statusPublished - 1 Jun 2016

Publication series

NameMethods in Molecular Biology
Volume1440
ISSN (Print)1064-3745

Fingerprint

Teichoic Acids
Cell Wall
Staphylococcus aureus
Peptidoglycan
Phosphates
Ribitol
Amino Sugars
Monosaccharides
Gram-Positive Bacteria
Polymerization
Glycerol
Anions
Chromatography
Polyacrylamide Gel Electrophoresis
Immune System
Hydrolysis
Bacteria
Amino Acids
Acids

Keywords

  • Bacterial cell surface
  • Bacterial cell wall
  • Bacterial glycopolymers
  • HPAEC-PAD
  • HPLC
  • Monosaccharide analysis
  • PAGE
  • Staphylococcus aureus
  • Wall teichoic acids (WTA)

Cite this

Covas, G., Vaz, F., Henriques, G., Pinho, M. G., & Filipe, S. R. (2016). Analysis of cell wall teichoic acids in Staphylococcus aureus. In Methods in Molecular Biology (Vol. 1440, pp. 201-213). (Methods in Molecular Biology; Vol. 1440). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-3676-2_15
Covas, Gonçalo ; Vaz, Filipa ; Henriques, Gabriela ; Pinho, Mariana G. ; Filipe, Sérgio R. / Analysis of cell wall teichoic acids in Staphylococcus aureus. Methods in Molecular Biology. Vol. 1440 Humana Press Inc., 2016. pp. 201-213 (Methods in Molecular Biology).
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Covas, G, Vaz, F, Henriques, G, Pinho, MG & Filipe, SR 2016, Analysis of cell wall teichoic acids in Staphylococcus aureus. in Methods in Molecular Biology. vol. 1440, Methods in Molecular Biology, vol. 1440, Humana Press Inc., pp. 201-213. https://doi.org/10.1007/978-1-4939-3676-2_15

Analysis of cell wall teichoic acids in Staphylococcus aureus. / Covas, Gonçalo; Vaz, Filipa; Henriques, Gabriela; Pinho, Mariana G.; Filipe, Sérgio R.

Methods in Molecular Biology. Vol. 1440 Humana Press Inc., 2016. p. 201-213 (Methods in Molecular Biology; Vol. 1440).

Research output: Chapter in Book/Report/Conference proceedingChapter

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Covas G, Vaz F, Henriques G, Pinho MG, Filipe SR. Analysis of cell wall teichoic acids in Staphylococcus aureus. In Methods in Molecular Biology. Vol. 1440. Humana Press Inc. 2016. p. 201-213. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-3676-2_15