Localization of Protein A on the Cell Surface of Staphylococcus aureus Cowan I and Protein A-Deficient Strains

1984 ◽  
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Surface ◽  
Protein A

scholarly journals Staphylococcal Surface Display of Immunoglobulin A (IgA)- and IgE-Specific In Vitro-Selected Binding Proteins (Affibodies) Based on Staphylococcus aureus Protein A

1999 ◽  
Vol 65 (9) ◽  
pp. 4134-4140 ◽  
Author(s):  
Elin Gunneriusson ◽  
Patrik Samuelson ◽  
Jenny Ringdahl ◽  
Hans Grönlund ◽  
Per-Åke Nygren ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Surface ◽  
Binding Capacity ◽  
Expression System ◽  
Surface Display ◽  
Protein A ◽  
Surface Proteins ◽  
Phage Library ◽  
Protein Chemistry ◽  
Ige Binding

ABSTRACT An expression system designed for cell surface display of hybrid proteins on Staphylococcus carnosus has been evaluated for the display of Staphylococcus aureus protein A (SpA) domains, normally binding to immunoglobulin G (IgG) Fc but here engineered by combinatorial protein chemistry to yield SpA domains, denoted affibodies, with new binding specificities. Such affibodies, with human IgA or IgE binding activity, have previously been selected from a phage library, based on an SpA domain. In this study, these affibodies have been genetically introduced in monomeric or dimeric forms into chimeric proteins expressed on the surface of S. carnosus by using translocation signals from aStaphylococcus hyicus lipase construct together with surface-anchoring regions of SpA. The recombinant surface proteins, containing the IgA- or IgE-specific affibodies, were demonstrated to be expressed as full-length proteins, localized and properly exposed at the cell surface of S. carnosus. Furthermore, these chimeric receptors were found to be functional, since recombinantS. carnosus cells were shown to have gained IgA and IgE binding capacity, respectively. In addition, a positive effect in terms of IgA and IgE reactivity was observed when dimeric versions of the affibodies were present. Potential applications for recombinant bacteria with redirected binding specificity in their surface proteins are discussed.

scholarly journals Staphylococcus aureus Fibronectin-Binding Protein A Mediates Cell-Cell Adhesion through Low-Affinity Homophilic Bonds

mBio ◽  
2015 ◽  
Vol 6 (3) ◽  
Author(s):  
Philippe Herman-Bausier ◽  
Sofiane El-Kirat-Chatel ◽  
Timothy J. Foster ◽  
Joan A. Geoghegan ◽  
Yves F. Dufrêne
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Surface ◽  
Biofilm Formation ◽  
Protein A ◽  
Intercellular Adhesion ◽  
Surface Proteins ◽  
Content Type ◽  
Fibronectin Binding ◽  
A Domains ◽  
Cell Cell

ABSTRACT Staphylococcus aureus is an important opportunistic pathogen which is a leading cause of biofilm-associated infections on indwelling medical devices. The cell surface-located fibronectin-binding protein A (FnBPA) plays an important role in the accumulation phase of biofilm formation by methicillin-resistant S. aureus (MRSA), but the underlying molecular interactions are not yet established. Here, we use single-cell and single-molecule atomic force microscopy to unravel the mechanism by which FnBPA mediates intercellular adhesion. We show that FnBPA is responsible for specific cell-cell interactions that involve the FnBPA A domain and cause microscale cell aggregation. We demonstrate that the strength of FnBPA-mediated adhesion originates from multiple low-affinity homophilic interactions between FnBPA A domains on neighboring cells. Low-affinity binding by means of FnBPA may be important for biofilm dynamics. These results provide a molecular basis for the ability of FnBPA to promote cell accumulation during S. aureus biofilm formation. We speculate that homophilic interactions may represent a generic strategy among staphylococcal cell surface proteins for guiding intercellular adhesion. As biofilm formation by MRSA strains depends on proteins rather than polysaccharides, our approach offers exciting prospects for the design of drugs or vaccines to inhibit protein-dependent intercellular interactions in MRSA biofilms. IMPORTANCE Staphylococcus aureus is a human pathogen that forms biofilms on indwelling medical devices, such as central venous catheters and prosthetic joints. This leads to biofilm infections that are difficult to treat with antibiotics because many cells within the biofilm matrix are dormant. The fibronectin-binding proteins (FnBPs) FnBPA and FnBPB promote biofilm formation by clinically relevant methicillin-resistant S. aureus (MRSA) strains, but the molecular mechanisms involved remain poorly understood. We used atomic force microscopy techniques to demonstrate that FnBPA mediates cell-cell adhesion via multiple, low-affinity homophilic bonds between FnBPA A domains on adjacent cells. Therefore, FnBP-mediated homophilic interactions represent an interesting target to prevent MRSA biofilms. We propose that such homophilic mechanisms may be widespread among staphylococcal cell surface proteins, providing a means to guide intercellular adhesion and biofilm accumulation.

Effects of fibronectin on the compact colony formation in staphylococci

1984 ◽  
Vol 30 (3) ◽  
pp. 419-422 ◽  
Author(s):  
Yukio Usui ◽  
Yukio Ohshima ◽  
Kosaku Yoshida
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Surface ◽  
Staphylococcus Epidermidis ◽  
Colony Formation ◽  
Protein A ◽  
Soft Agar ◽  
Cellular Factor ◽  
Diffuse Growth

Fifty-two unencapsulated strains of Staphylococcus aureus, including strains of Wood 46 and Cowan I, formed compact colonies in fibronectin – soft agar. However, 20 encapsulated strains of Staphylococcus aureus and 50 strains of Staphylococcus epidermidis showed diffuse growth in the medium. The results suggest that another possible cellular factor, other than protein A, is involved in the binding of the cell surface with fibronectin and that it would be one of factors in forming compact colonies in serum – soft agar.

scholarly journals Regulation of agr-Dependent Virulence Genes in Staphylococcus aureus by RNAIII from Coagulase-Negative Staphylococci

1998 ◽  
Vol 180 (12) ◽  
pp. 3181-3186 ◽  
Author(s):  
Karin Tegmark ◽  
Eva Morfeldt ◽  
Staffan Arvidson
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Surface ◽  
Serine Protease ◽  
Protein A ◽  
Surface Proteins ◽  
Open Reading Frames ◽  
Alpha Toxin ◽  
Coagulase Negative Staphylococci ◽  
Cell Surface Proteins ◽  
Genes Encoding

ABSTRACT Many of the genes coding for extracellular toxins, enzymes, and cell surface proteins in Staphylococcus aureus are regulated by a 510-nucleotide (nt) RNA molecule, RNAIII. Transcription of genes encoding secreted toxins and enzymes, includinghla (alpha-toxin), saeB (enterotoxin B),tst (toxic shock syndrome toxin 1), and ssp(serine protease), is stimulated, while transcription of genes encoding cell surface proteins, like spa (protein A) andfnb (fibronectin binding proteins), is repressed. Besides being a regulator, RNAIII is also an mRNA coding for staphylococcal delta-lysin. We have identified RNAIII homologs in three different coagulase-negative staphylococci (CoNS), i.e., Staphylococcus epidermidis, Staphylococcus simulans, andStaphylococcus warneri. RNAIII from these CoNS turned out to be very similar to that of S. aureus and contained open reading frames encoding delta-lysin homologs. Though a number of big insertions and/or deletions have occurred, mainly in the 5′ half of the molecules, the sequences show a high degree of identity, especially in the first 50 and last 150 nt. The CoNS RNAIII had the ability to completely repress transcription of protein A in an RNAIII-deficientS. aureus mutant and the ability to stimulate transcription of the alpha-toxin and serine protease genes. However, the stimulatory effect was impaired compared to that of S. aureus RNAIII, suggesting that these regulatory functions are independent. By creating S. epidermidis-S. aureus RNAIII hybrids, we could also show that both the 5′ and 3′ halves of the RNAIII molecule are involved in the transcriptional regulation of alpha-toxin and serine protease mRNAs in S. aureus.

scholarly journals Staphylococcus aureusProtein A Mediates Interspecies Interactions at the Cell Surface ofPseudomonas aeruginosa

mBio ◽  
2016 ◽  
Vol 7 (3) ◽  
Author(s):  
Catherine R. Armbruster ◽  
Daniel J. Wolter ◽  
Meenu Mishra ◽  
Hillary S. Hayden ◽  
Matthew C. Radey ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Microbial Community ◽  
Cell Surface ◽  
Biofilm Formation ◽  
Protein A ◽  
Staphylococcal Protein A ◽  
Interspecies Interactions ◽  
Community Interactions ◽  
Staphylococcal Protein

ABSTRACTWhile considerable research has focused on the properties of individual bacteria, relatively little is known about how microbial interspecies interactions alter bacterial behaviors and pathogenesis.Staphylococcus aureusfrequently coinfects with other pathogens in a range of different infectious diseases. For example, coinfection byS. aureuswithPseudomonas aeruginosaoccurs commonly in people with cystic fibrosis and is associated with higher lung disease morbidity and mortality.S. aureussecretes numerous exoproducts that are known to interact with host tissues, influencing inflammatory responses. The abundantly secretedS. aureusstaphylococcal protein A (SpA) binds a range of human glycoproteins, immunoglobulins, and other molecules, with diverse effects on the host, including inhibition of phagocytosis ofS. aureuscells. However, the potential effects of SpA and otherS. aureusexoproducts on coinfecting bacteria have not been explored. Here, we show thatS. aureus-secreted products, including SpA, significantly alter two behaviors associated with persistent infection. We found that SpA inhibited biofilm formation by specificP. aeruginosaclinical isolates, and it also inhibited phagocytosis by neutrophils of all isolates tested. Our results indicate that these effects were mediated by binding to at least twoP. aeruginosacell surface structures—type IV pili and the exopolysaccharide Psl—that confer attachment to surfaces and to other bacterial cells. Thus, we found that the role of a well-studiedS. aureusexoproduct, SpA, extends well beyond interactions with the host immune system. Secreted SpA alters multiple persistence-associated behaviors of another common microbial community member, likely influencing cocolonization and coinfection with other microbes.IMPORTANCEBacteria rarely exist in isolation, whether on human tissues or in the environment, and they frequently coinfect with other microbes. However, relatively little is known about how microbial interspecies interactions alter bacterial behaviors and pathogenesis. We identified a novel interaction between two bacterial species that frequently infect together—Staphylococcus aureusandPseudomonas aeruginosa. We show that theS. aureus-secreted protein staphylococcal protein A (SpA), which is well-known for interacting with host targets, also binds to specificP. aeruginosacell surface molecules and alters two persistence-associatedP. aeruginosabehaviors: biofilm formation and uptake by host immune cells. BecauseS. aureusfrequently precedesP. aeruginosain chronic infections, these findings reveal how microbial community interactions can impact persistence and host interactions during coinfections.

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