Clumping Factor B, a Fibrinogen-binding MSCRAMM (Microbial Surface Components Recognizing Adhesive Matrix Molecules) Adhesin of Staphylococcus aureus, Also Binds to the Tail Region of Type I Cytokeratin 10

Walsh, Evelyn J.; O’Brien, Louise; Liang, Xiaowen; Höök, Magnus; Foster, Timothy J.

doi:10.1074/jbc.m408713200

Cited by 107 publications

(106 citation statements)

References 31 publications

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“…4D). The interaction of S. aureus with cytokeratin 10, a major substituent of human skin, is mediated by at least two CWAassociated proteins, including ClfB and IsdA (5,54), and is considered to be important for S. aureus nasal colonization (56). The identification of a cytokeratin 10-binding surface protein of S. pseudintermedius suggests that such an interaction may also be important for canine host colonization.…”

Section: Vol 79 2011mentioning

confidence: 99%

“…In addition, fibrinogen was purified from canine, equine, and avian blood (Lampire) by modified cold ethanol fractionation according to previously published protocols (6,9). Recombinant cytokeratin 10 was expressed and purified from a pQE30 expression clone which encoded the C terminus of mouse cytokeratin 10 (rMCK10 294-570) (54). Recombinant protein expression in E. coli XL1-Blue cells (Stratagene, United Kingdom) was induced with IPTG (0.5 mM) and protein purified under hybrid conditions according to the manufacturer's instructions using a Ni-nitrilotriacetic acid (NTA) benchtop purification system (Invitrogen, United Kingdom).…”

Section: Methodsmentioning

confidence: 99%

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Genomic and Surface Proteomic Analysis of the Canine Pathogen Staphylococcus pseudintermedius Reveals Proteins That Mediate Adherence to the Extracellular Matrix

et al. 2011

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Cell wall-associated (CWA) proteins made by Gram-positive pathogens play a fundamental role in pathogenesis. Staphylococcus pseudintermedius is a major animal pathogen responsible for the canine skin disease bacterial pyoderma. Here, we describe the bioinformatic analysis of the family of 18 predicted CWA proteins encoded in the genome of S. pseudintermedius strain ED99 and determine their distribution among a phylogenetically diverse panel of S. pseudintermedius clinical isolates and closely related species of the Staphylococcus intermedius group. In parallel, we employed a proteomic approach to identify proteins presented on the surface of strain ED99 in vitro, revealing a total of 60 surface-localized proteins in one or more phases of growth, including 6 of the 18 genome-predicted CWA proteins. Based on these analyses, we selected two CWA proteins (SpsD and SpsL) encoded by all strains examined and investigated their capacity to mediate adherence to extracellular matrix proteins. We discovered that SpsD and SpsL mediated binding of a heterologous host, Lactococcus lactis, to fibrinogen and fibronectin and that SpsD mediated binding to cytokeratin 10, a major constituent of mammalian skin. Of note, the interaction with fibrinogen was host-species dependent, suggestive of a role for SpsD and SpsL in the host tropism of S. pseudintermedius. Finally, we identified IgG specific for SpsD and SpsL in sera from dogs with bacterial pyoderma, implying that both proteins are expressed during infection. The combined genomic and proteomic approach employed in the current study has revealed novel host-pathogen interactions which represent candidate therapeutic targets for the control of bacterial pyoderma.

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Section: Vol 79 2011mentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Genomic and Surface Proteomic Analysis of the Canine Pathogen Staphylococcus pseudintermedius Reveals Proteins That Mediate Adherence to the Extracellular Matrix

et al. 2011

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“…Binding inhibition studies were performed in microtiter plates coated with recombinant mouse cytokeratin 10 (1 g/well) expressed and purified as previously described (49). A suspension containing 10 8 CFU S. aureus Newman was incubated for 1 h at 22°C with increasing amounts of MAbs before the bacteria were added to the coated microtiter wells.…”

Section: Methodsmentioning

confidence: 99%

Immunization with Staphylococcus aureus Clumping Factor B, a Major Determinant in Nasal Carriage, Reduces Nasal Colonization in a Murine Model

et al. 2006

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Staphylococcus aureus is responsible for a wide range of infections, including soft tissue infections and potentially fatal bacteremias. The primary niche for S. aureus in humans is the nares, and nasal carriage is a documented risk factor for staphylococcal infection. Previous studies with rodent models of nasal colonization have implicated capsule and teichoic acid as staphylococcal surface factors that promote colonization. In this study, a mouse model of nasal colonization was utilized to demonstrate that S. aureus mutants that lack clumping factor A, collagen binding protein, Staphylococcus aureus causes a diverse spectrum of severe infections in humans, including bacteremia, endocarditis, and osteomyelitis, as well as skin and soft tissue infections. Notorious for decades as a major source of nosocomial infections, S. aureus has recently taken on a new role in causing an escalating number of community-acquired infections. To offset the problems of antibiotic-resistant S. aureus strains, preventive measures (e.g., immunization) should be explored as a complement to existing therapeutic approaches aimed at controlling this bacterial pathogen.Humans are a reservoir for S. aureus, and the nose is the principal site of staphylococcal colonization. Approximately 20% of people persistently carry S. aureus in the anterior nares, ϳ60% are intermittent carriers, and ϳ20% are noncarriers (19). Nasal carriage is a known risk factor for staphylococcal infection in a number of clinical settings (51). Certain patient populations that show higher rates of S. aureus nasal colonization have an increased risk of staphylococcal infection. These populations include patients with diabetes, eczema, and human immunodeficiency virus infection, individuals receiving continuous ambulatory peritoneal dialysis or hemodialysis, and injection drug users (19). Moreover, patients in hospitals or individuals living in crowded conditions often show higherthan-normal rates of S. aureus nasal colonization. The source of ϳ80% of S. aureus bacteremias is endogenous since infecting bacteria have been shown by genotypic analysis to be identical to organisms recovered from the nasal mucosa (48, 53). These observations support an approach in which systemic S. aureus infections are prevented by eliminating or reducing nasal carriage.One approach commonly used to reduce S. aureus carriage in individuals at risk for staphylococcal infection involves topical treatment with a nasal ointment containing the antibiotic mupirocin. Eradication of nasal carriage with topical mupirocin has been correlated with a reduction in the incidence of S. aureus infection in some patient populations (20, 45), but not in others (40,54). Whereas mupirocin is effective in decolonizing nasal carriers, recolonization often occurs from extranasal carriage sites (52). Of further concern is the emergence of mupirocin resistance in S. aureus (31, 46). The utility of

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“…The clumping factor B gene (clfB) of S. aureus encodes an MSCRAMM protein that binds to fibrinogen (30) and keratin and facilitates S. aureus colonization in human nares (32,53). Clumping factor B has also been implicated in the pathogenesis of S. aureusinduced endocarditis (11).…”

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confidence: 99%

Comparative Sequencing of the Serine-Aspartate Repeat-Encoding Region of the Clumping Factor B Gene ( clfB ) for Resolution within Clonal Groups of Staphylococcus aureus

Koreen

Ramaswamy

Naidich³

et al. 2005

J Clin Microbiol

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Molecular techniques such as spa typing and multilocus sequence typing use DNA sequence data for differentiating Staphylococcus aureus isolates. Although spa typing is capable of detecting both genetic microand macrovariation, it has less discriminatory power than the more labor-intensive pulsed-field gel electrophoresis (PFGE) and costly genomic DNA microarray analyses. This limitation hinders strain interrogation for newly emerging clones and outbreak investigations in hospital or community settings where robust clones are endemic. To overcome this constraint, we developed a typing system using DNA sequence analysis of the serine-aspartate (SD) repeat-encoding region within the gene encoding the keratin-and fibrinogen-binding clumping factor B (clfB typing) and tested whether it is capable of discriminating within clonal groups. We analyzed 116 S. aureus strains, and the repeat region was present in all isolates, varying in sequence and in length from 420 to 804 bp. In a sample of 36 well-characterized genetically diverse isolates, clfB typing subdivided identical spa and PFGE clusters which had been discriminated by whole-genome DNA microarray mapping. The combination of spa typing and clfB typing resulted in a discriminatory power (99.5%) substantially higher than that of spa typing alone and closely approached that of the whole-genome microarray (100.0%). clfB typing also successfully resolved genetic differences among isolates differentiated by PFGE that had been collected over short periods of time from single hospitals and that belonged to the most prevalent S. aureus clone in the United States. clfB typing demonstrated in vivo, in vitro, and interpatient transmission stability yet revealed that this locus may be recombinogenic in a primarily clonal population structure. Taken together, these data show that the SD repeat-encoding region of clfB is a highly stable marker of microvariation, that in conjunction with spa typing it may serve as a DNA sequence-based alternative to PFGE for investigating genetically similar strains, and that it is useful for analyzing collections of isolates in both long-term population-based and local epidemiologic studies.Strain typing techniques for Staphylococcus aureus, the leading cause of nosocomial infections (28), have become widely used. These techniques aid in both local/short-term epidemiologic outbreak investigations and global/long-term population-based studies of methicillin-resistant and -susceptible S. aureus (MRSA and MSSA, respectively). Macrorestriction digests using pulsed-field gel electrophoresis (PFGE) have been shown to be highly effective in outbreak settings (48). Multilocus enzyme electrophoresis (MLEE), multilocus sequence typing (MLST), and sequence analysis of the repeat region within the coagulase gene, i.e., coa typing, are effective techniques for analyzing S. aureus strains in long-term study settings (10,42), and the sequence analysis of the repeat region within the protein A gene, spa typing, can effectively be used in both settings (22,41)....

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Clumping Factor B, a Fibrinogen-binding MSCRAMM (Microbial Surface Components Recognizing Adhesive Matrix Molecules) Adhesin of Staphylococcus aureus, Also Binds to the Tail Region of Type I Cytokeratin 10

Cited by 107 publications

References 31 publications

Genomic and Surface Proteomic Analysis of the Canine Pathogen Staphylococcus pseudintermedius Reveals Proteins That Mediate Adherence to the Extracellular Matrix

Genomic and Surface Proteomic Analysis of the Canine Pathogen Staphylococcus pseudintermedius Reveals Proteins That Mediate Adherence to the Extracellular Matrix

Immunization with Staphylococcus aureus Clumping Factor B, a Major Determinant in Nasal Carriage, Reduces Nasal Colonization in a Murine Model

Comparative Sequencing of the Serine-Aspartate Repeat-Encoding Region of the Clumping Factor B Gene ( clfB ) for Resolution within Clonal Groups of Staphylococcus aureus

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