Analysis of Cell Wall Teichoic Acids in Staphylococcus aureus.
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).
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
G
Covas
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.
Filipa
F
Vaz
Laboratory of Bacterial Cell Biology, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal.
Gabriela
G
Henriques
Laboratory of Bacterial Cell Biology, Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal.
Mariana G
MG
Pinho
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. sfilipe@itqb.unl.pt.|UCIBIO@REQUIMTE, Departamento de Ciências da Vida/ Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Campus da Caparica, 2829-516, Caparica, Portugal. sfilipe@itqb.unl.pt.
Sérgio R
SR
Filipe
D001426:Bacterial Proteins / Q000378:metabolism; D002473:Cell Wall / Q000378:metabolism; D002852:Chromatography, Ion Exchange; D006868:Hydrolysis; D010710:Phosphates / Q000737:chemistry; D013211:Staphylococcus aureus / Q000378:metabolism*; D013682:Teichoic Acids / Q000032:analysis* / Q000737:chemistry
D001426:Bacterial Proteins; D010710:Phosphates; D013682:Teichoic Acids
Bacterial cell surface; Bacterial cell wall; Bacterial glycopolymers; HPAEC-PAD; HPLC; Monosaccharide analysis; PAGE; Staphylococcus aureus; Wall teichoic acids (WTA)