Isolation, structural characterization, and immunological evaluation of a high-molecular-weight exopolysaccharide from Staphylococcus aureus

Joyce, Joseph G.; Abeygunawardana, Chitrananda; Xu, Qiuwei; Cook, James C.; Hepler, Robert; Przysiecki, Craig T.; Grimm, Karen M.; Roper, Keith; Ip, Charlotte C. Yu; Cope, Leslie D.; Montgomery, Donna L.; Chang, Mason; Campie, Sherilyn; Brown, Martha; McNeely, Tessie; Zorman, Julie; Maira-Litrán, Tomas; Pier, Gerald B.; Keller, Paul M.; Jansen, Kathrin U.; Mark, George E.

doi:10.1016/s0008-6215(03)00045-4

Cited by 97 publications

(97 citation statements)

References 46 publications

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“…In comparison to other known exopolysaccharides that are neutral, negatively charged or, on occasion, zwitterionic with both positive and negative charges (Cobb et al, 2004;Cobb & Kasper, 2005), PNAG represents a very unusual bacterial exopolysaccharide molecule, as some GlcNAc residues become deacetylated by the IcaB protein (Vuong et al, 2004), which provides a positive net charge to the polymer (Joyce et al, 2003). The deacetylation process, and therefore the cationic character of PNAG, has been shown to be crucial for the attachment of PNAG to the negatively charged bacterial surface and the biofilm formation process in both S. epidermidis (Vuong et al, 2004) and S. aureus (Cerca et al, 2007).…”

Section: Discussionmentioning

confidence: 99%

“…aureus biofilm formation is generally mediated by the production of the extracellular polysaccharide adhesin termed both polysaccharide intercellular adhesin (PIA) or poly-N-acetylglucosamine (PNAG), whose synthesis depends on the expression of the biosynthetic enzymes encoded by the icaADBC operon (Cramton et al, 1999;Heilmann et al, 1996b;Mckenney et al, 1999). The exopolysaccharide PNAG is a linear b-(1-6)-linked Nacetylglucosaminoglycan with a high proportion of amino groups substituted with N-linked acetate, and a small amount of the hydroxyl groups esterified with acetate and succinate (Joyce et al, 2003;Mack et al, 1996;MairaLitrán et al, 2002). It has been shown recently that the IcaB protein is involved in the production of positive charge in the PNAG polymer by deacetylation of a proportion of the N-acetylglucosamines (GlcNAcs).…”

Section: Introductionmentioning

confidence: 99%

“…In favour of this hypothesis, it has been shown that TAs can complex with different polysaccharides in a relatively non-specific way (Doyle et al, 1975). In addition, TAs are usually found as contaminants in PNAG purification processes, although no covalent linkage between them has been found (Joyce et al, 2003;MairaLitrán et al, 2002;McKenney et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

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Wall teichoic acids are dispensable for anchoring the PNAG exopolysaccharide to the Staphylococcus aureus cell surface

et al. 2008

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Biofilm formation in Staphylococcus aureus is usually associated with the production of the poly-N-acetylglucosamine (PNAG) exopolysaccharide, synthesized by proteins encoded by the icaADBC operon. PNAG is a linear b-(1-6)-linked N-acetylglucosaminoglycan that has to be partially deacetylated and consequently positively charged in order to be associated with bacterial cell surfaces. Here, we investigated whether attachment of PNAG to bacterial surfaces is mediated by ionic interactions with the negative charge of wall teichoic acids (WTAs), which represent the most abundant polyanions of the Gram-positive bacterial envelope. We generated WTA-deficient mutants by in-frame deletion of the tagO gene in two genetically unrelated S. aureus strains. The DtagO mutants were more sensitive to high temperatures, showed a higher degree of cell aggregation, had reduced initial adherence to abiotic surfaces and had a reduced capacity to form biofilms under both steady-state and flow conditions. However, the levels as well as the strength of the PNAG interaction with the bacterial cell surface were similar between DtagO mutants and their corresponding wild-type strains. Furthermore, double DtagO DicaADBC mutants displayed a similar aggregative phenotype to that of single DtagO mutants, indicating that PNAG is not responsible for the aggregative behaviour observed in DtagO mutants. Overall, the absence of WTAs in S. aureus had little effect on PNAG production or anchoring to the cell surface, but did affect the biofilm-forming capacity, cell aggregative behaviour and the temperature sensitivity/stability of S. aureus.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Wall teichoic acids are dispensable for anchoring the PNAG exopolysaccharide to the Staphylococcus aureus cell surface

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“…cinyl moieties were later determined to be artifacts of purification (29). Poly-␤-1,6-GlcNAc was subsequently isolated from Escherichia coli and shown to depend on the pgaABCD operon for its production (50).…”

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confidence: 99%

Roles of pgaABCD Genes in Synthesis, Modification, and Export of the Escherichia coli Biofilm Adhesin Poly-β-1,6- N -Acetyl- d -Glucosamine

et al. 2008

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The linear homopolymer poly-␤-1,6-N-acetyl-D-glucosamine (␤-1,6-GlcNAc; PGA) serves as an adhesin for the maintenance of biofilm structural stability in diverse eubacteria. Its function in Escherichia coli K-12 requires the gene products of the pgaABCD operon, all of which are necessary for biofilm formation. PgaC is an apparent glycosyltransferase that is required for PGA synthesis. Using a monoclonal antibody directed against E. coli PGA, we now demonstrate that PgaD is also needed for PGA formation. The deletion of genes for the predicted outer membrane proteins PgaA and PgaB did not prevent PGA synthesis but did block its export, as shown by the results of immunoelectron microscopy (IEM) and antibody adsorption assays. IEM also revealed a conditional localization of PGA at the cell poles, the initial attachment site for biofilm formation. PgaA contains a predicted ␤-barrel porin and a superhelical domain containing tetratricopeptide repeats, which may mediate protein-protein interactions, implying that it forms the outer membrane secretin for PGA. PgaB contains predicted carbohydrate binding and polysaccharide N-deacetylase domains. The overexpression of pgaB increased the primary amine content (glucosamine) of PGA. Site-directed mutations targeting the N-deacetylase catalytic activity of PgaB blocked PGA export and biofilm formation, implying that N-deacetylation promotes PGA export through the PgaA porin. The results of previous studies indicated that N-deacetylation of ␤-1,6-GlcNAc in Staphylococcus epidermidis by the PgaB homolog, IcaB, anchors it to the cell surface. The deletion of icaB resulted in release of ␤-1,6-GlcNAc into the growth medium. Thus, covalent modification of ␤-1,6-GlcNAc by N-deacetylation serves distinct biological functions in gram-negative and gram-positive species, dictated by cell envelope differences.

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“…Production of staphylococcal ␤-1,6-GlcNAc polysaccharides, alternatively known as PIA, PS/A, SAE, and PNAG (14,17,22,23,25,26), depends upon the icaABCD locus. Two gene products of the pga and ica operons are apparently homologous (37).…”

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confidence: 99%

Depolymerization of β-1,6-N-Acetyl-d-Glucosamine Disrupts the Integrity of Diverse Bacterial Biofilms

et al. 2005

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Polymeric ␤-1,6-N-acetyl-D-glucosamine (poly-␤-1,6-GlcNAc) has been implicated as an Escherichia coli and Staphylococcus epidermidis biofilm adhesin, the formation of which requires the pgaABCD and icaABCD loci, respectively. Enzymatic hydrolysis of poly-␤-1,6-GlcNAc, demonstrated for the first time by chromatography and mass spectrometry, disrupts biofilm formation by these species and by Yersinia pestis and Pseudomonas fluorescens, which possess pgaABCD homologues.

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Isolation, structural characterization, and immunological evaluation of a high-molecular-weight exopolysaccharide from Staphylococcus aureus

Cited by 97 publications

References 46 publications

Wall teichoic acids are dispensable for anchoring the PNAG exopolysaccharide to the Staphylococcus aureus cell surface

Wall teichoic acids are dispensable for anchoring the PNAG exopolysaccharide to the Staphylococcus aureus cell surface

Roles of pgaABCD Genes in Synthesis, Modification, and Export of the Escherichia coli Biofilm Adhesin Poly-β-1,6- N -Acetyl- d -Glucosamine

Depolymerization of β-1,6-N-Acetyl-d-Glucosamine Disrupts the Integrity of Diverse Bacterial Biofilms

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