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Analysis of Cell Wall Teichoic Acids in Staphylococcus aureus

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Bacterial Cell Wall Homeostasis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1440))

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).

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Acknowledgements

This work was supported by PhD fellowships from the Portuguese Science Agency (FCT): SFRH/BD/52207/2013 (GC) and SFRH/BD/78748/2011 (FV). Research work described was also supported by grants PTDC/BIA-PLA/3432/2012 and IF/01464/2013 (SF).

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Authors and Affiliations

  1. Laboratory of Bacterial Cell Surfaces and Pathogenesis, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Avenida da República, Apartado 127, 2781-901, Oeiras, Portugal

    Gonçalo Covas, Filipa Vaz & Sérgio R. Filipe

  2. Laboratory of Bacterial Cell Biology, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal

    Gabriela Henriques & Mariana G. Pinho

  3. UCIBIO@REQUIMTE, Departamento de Ciências da Vida/ Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Campus da Caparica, 2829-516, Caparica, Portugal

    Sérgio R. Filipe

Authors
  1. Gonçalo Covas
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  2. Filipa Vaz
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  3. Gabriela Henriques
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  4. Mariana G. Pinho
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  5. Sérgio R. Filipe
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Corresponding author

Correspondence to Sérgio R. Filipe .

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Editors and Affiliations

  1. Dept. of Biochem & Dept. of Bio and Med., Univ. of Cambridge andOxford Brookes Unv, Cambridge, United Kingdom

    Hee-Jeon Hong

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Covas, G., Vaz, F., Henriques, G., Pinho, M.G., Filipe, S.R. (2016). Analysis of Cell Wall Teichoic Acids in Staphylococcus aureus . In: Hong, HJ. (eds) Bacterial Cell Wall Homeostasis. Methods in Molecular Biology, vol 1440. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3676-2_15

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  • DOI: https://doi.org/10.1007/978-1-4939-3676-2_15

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3674-8

  • Online ISBN: 978-1-4939-3676-2

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