Sorting of protein a to the staphylococcal cell wall

Schneewind, Olaf; Model, Peter; Fischetti, Vincent A.

doi:10.1016/0092-8674(92)90101-h

Cited by 505 publications

(499 citation statements)

References 80 publications

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“…At the Cterminal end of the S. aureus MSCRAMMs are features required for cell-wall anchoring, including an LPXTG (X being any amino acid residue) motif preceded by a cellwall-spanning domain, followed by a hydrophobic transmembrane region and, finally, a cytosolic tail composed of a short sequence rich in positively charged amino acid residues (Patti & Hook, 1994). The LPXTG motif is recognized by a transpeptidase called sortase which is responsible for catalysing the covalent linking of these proteins to the staphylococcal cell wall (Mazmanian et al, 2001;Schneewind et al, 1992). This cell-wall anchoring mechanism seems to be widely utilized among Grampositive bacteria, since sortase homologues have been identified in almost all Gram-positive species examined (Pallen et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

A family of putative MSCRAMMs from Enterococcus faecalis

et al. 2004

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The recently published Enterococcus faecalis genome [Paulsen, I. T., Banerjei, L., Myers, G. S. & 29 other authors (2003). Science 299, 2071–2074)] was examined and 41 putative cell-wall-anchored proteins were identified. Seventeen of these proteins are predicted to contain tandemly repeated immunoglobulin-like folds characteristic of the structural organization of staphylococcal adhesins of the MSCRAMM (microbial surface component recognizing adhesive matrix molecules) type. Two of the nine proteins selected for further study appear to represent cell-wall-anchored enzymes. It is proposed that the remaining seven proteins constitute a family of structurally related proteins potentially interacting with proteins of the host. This family includes the previously identified collagen/laminin-binding MSCRAMM ACE [Rich, R. L., Kreikemeyer, B., Owens, R. T., LaBrenz, S., Narayana, S. V., Weinstock, G. M., Murray, B. E. & Hook, M. (1999). J Biol Chem 274, 26939–26945]. It is further demonstrated that genes encoding the seven putative MSCRAMMs are present in all E. faecalis strains tested and these proteins appear to be expressed during infection in humans, since sera from infected individuals contain antibodies reacting with recombinant versions of the enterococcal proteins.

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Section: Introductionmentioning

confidence: 99%

A family of putative MSCRAMMs from Enterococcus faecalis

et al. 2004

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“…SrtA is responsible for anchoring proteins containing a C-terminal tripartite sorting signal, which consists of 1) an LPXTG pentapeptide followed by 2) a less well conserved hydrophobic domain and 3) a basic charged tail (18,19). Both the hydrophobic domain and the charged tail help to retain the putative surface protein in the membrane prior to sortase-catalyzed anchoring.…”

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confidence: 99%

Engineering the Substrate Specificity of Staphylococcus aureus Sortase A

Bentley

Gaweska

Kielec

et al. 2007

Journal of Biological Chemistry

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The Staphylococcus aureus transpeptidase Sortase A (SrtA) anchors virulence and colonization-associated surface proteins to the cell wall. SrtA selectively recognizes a C-terminal LPXTG motif, whereas the related transpeptidase Sortase B (SrtB) recognizes a C-terminal NPQTN motif. In both enzymes, cleavage occurs after the conserved threonine, followed by amide bond formation between threonine and the pentaglycine cross-bridge of cell wall peptidoglycan. Genetic and biochemical studies strongly suggest that SrtA and SrtB exhibit exquisite specificity for their recognition motifs. To better understand the origins of substrate specificity within these two isoforms, we used sequence and structural analysis to predict residues and domains likely to be involved in conferring substrate specificity. Mutational analyses and domain swapping experiments were conducted to test their function in substrate recognition and specificity. Marked changes in the specificity profile of SrtA were obtained by replacing the ␤6/␤7 loop in SrtA with the corresponding domain from SrtB. The chimeric ␤6/␤7 loop swap enzyme (SrtLS) conferred the ability to acylate NPQTNcontaining substrates, with a k cat /K m app of 0.0062 ؎ 0.003 M ؊1 s ؊1 . This enzyme was unable to perform the transpeptidation stage of the reaction, suggesting that additional domains are required for transpeptidation to occur. The overall catalytic specificity profile (k cat /K m app (NPQTN)/k cat /K m app (LPETG)) of SrtLS was altered 700,000-fold from SrtA. These results indicate that the ␤6/␤7 loop is an important site for substrate recognition in sortases.

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“…This is likely rooted in the ability of the bacteria to evade and/or destroy important components of their host defenses [10,11]. Protein A is anchored on the peptidoglycan cell wall and is also secreted during the exponential growth phase [12–15]. It is an important virulence factor, able to bind IgG via its Fc region [16–18] protecting staphylococci from opsonophagocytosis.…”

Section: Introductionmentioning

confidence: 99%

Characterization of recombinant wild-type and nontoxigenic protein A from Staphylococcus pseudintermedius

2018

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Background: Staphylococcus pseudintermedius is an opportunistic pathogen that is the major cause of pyoderma affecting dogs. Conventional antimicrobial treatment for infections caused by this organism have failed in recent years due to widespread resistance and alternative treatment strategies are a high priority. Protein A encoded in Staphylococcus aureus by spa protects the bacterium by binding IgG and acts as a superantigen. Staphylococcus pseudintermedius possess two genes orthologous to S. aureus spa, spsP, and spsQ. Methods: SpsQ and SpsQ-M, a non-toxigenic SpsQ, were cloned and expressed as recombinant proteins and their cytotoxic effect on canine B cells was measured. The neutralizing ability of antibody raised against them in clinically healthy dogs was evaluated. Results: S. pseudintermedius SpsQ induced apoptosis of canine B cells. Specific amino acid substitutions diminished SpsQ-M binding to immunoglobulin and its super-antigenic activity, while its antigenicity was maintained. This recombinant, non-toxigenic S. pseudintermedius SpsQ stimulated the production of antibodies in dogs that specifically reacted with SpsQ and greatly diminished its cytotoxic effect on canine B cells. Conclusions: The production of neutralizing antibody suggests that attenuated, non-toxic SpsQ produced in this study is a good candidate for inclusion in a vaccine for use in the treatment and prevention of S. pseudintermedius infections.Abbreviations: SpA: Staphylococcus aureus protein A; SpsP: Staphylococcus pseudintermedius protein A; SpsQ: Staphylococcus pseudintermedius protein A; SpsQ-M: attenuated Staphylococcus pseudintermedius protein A; MRSP: methicillin resistant Staphylococcus pseudintermedius; IgA: immunoglobulin A; IgG: immunoglobulin G; IgM: immunoglobulin M; VH: variable region of immunoglobulin heavy chain; IgBD: immunoglobulin binding domains; MFI: mean fluorescent intensity; SEM: standard error of the mean; PBMC: Peripheral blood mononuclear cells; CD21: complement receptor type 2; ST: Sequence type; OD: Optical density; ORF: open reading frame; PBS: Phosphate buffered saline; Tween 20: Polyethylene glycol sorbitan monolaurate 20; HRP: horseradish peroxidase; TMB- 3,3',5,5'-Tetramethylbenzidine

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Sorting of protein a to the staphylococcal cell wall

Cited by 505 publications

References 80 publications

A family of putative MSCRAMMs from Enterococcus faecalis

A family of putative MSCRAMMs from Enterococcus faecalis

Engineering the Substrate Specificity of Staphylococcus aureus Sortase A

Characterization of recombinant wild-type and nontoxigenic protein A from Staphylococcus pseudintermedius

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