Identification of a Fibronectin-Binding Protein from Staphylococcus epidermidis

Abstract

Staphylococcus epidermidis has been reported to bind to a number of host cell extracellular matrix proteins, including fibronectin. Here we report the identification of a fibronectin-binding protein from S. epidermidis. A phage display library of S. epidermidis genomic DNA was constructed and panned against immobilized fibronectin. A number of phagemid clones containing overlapping inserts were identified, and one of these clones, pSE109FN, contained a 1.4-kb insert. Phage pSE109FN was found to bind to fibronectin but not to collagen, fibrinogen, laminin, or vitronectin. However, pSE109FN also bound to heparin, hyaluronate, and plasminogen, although to a lesser extent than it bound to fibronectin. Staphylococcus epidermidis, a normal skin commensal, is an important opportunistic pathogen responsible for infections of implanted medical devices and infections at sites of surgery (24). Medical devices infected by S. epidermidis include cardiac pacemakers (1), cerebrospinal fluid shunts (23), continuous ambulatory peritoneal dialysis catheters (14), intravascular catheters (2), orthopedic devices (11), and prosthetic heart valves (13). The accepted paradigm for S. epidermidis adhesion to biomaterial surfaces is a two-step process involving a primary attachment step mediated by hydrophobic interactions, followed by biofilm formation and intercellular interactions (6). However, it is acknowledged that the primary attachment step is mediated not only by hydrophobic interactions but also by bacterial proteins.Since foreign bodies become rapidly coated with plasma proteins when they are implanted in the body, S. epidermidis may bind to implants through interactions with immobilized extracellular matrix proteins. In vitro studies have shown that S. epidermidis can bind to biomaterials coated with fibronectin and fibrinogen (3,5,9,17). The ability of Staphylococcus aureus to bind to extracellular matrix proteins through a family of proteins termed the microbial surface components recognizing adhesive matrix molecules is well documented (for a review see reference 4). However, comparatively little is known about how S. epidermidis interacts with matrix proteins. It has been established that S. epidermidis binds to fibrinogen through SdrG, a microbial surface component recognizing adhesive matrix molecules, which is also known as Fbe (7,16,18,19). The autolysin AtlE of S. epidermidis, which has been implicated as the mediator of primary attachment of this organism to polystyrene surfaces, has also been shown to bind vitronectin (8). Although it has been demonstrated that S. epidermidis binds to fibronectin in vitro (3,5,9), no fibronectin-binding protein has been identified yet. In fact, it has been suggested that teichoic acid may mediate binding of S. epidermidis to immobilized fibronectin (10). The aim of the work described in this paper was to use shotgun phage display cloning to identify S. epidermidis genes that code for fibronectin-binding proteins. MATERIALS AND METHODSBacterial strains and plasmids. The type strain ...

Antibodies to PhnD Inhibit Staphylococcal Biofilms

“…The fluorescent reporter was then used to compare PhnD antibodies to those for other targets, including several proteins with a known role in S. epidermidis biofilm formation: EmbP (SERP1011), Aap (SERP2398), and SesC (SERP2264) (17)(18)(19)(20). Treatment with EmbP antibodies also led to a large decrease in biofilm formation (91% reduction versus the control); however, none of the other antibodies showed substantial effects at the endpoint (Fig.…”

The altered landscape of the human skin microbiome in patients with primary immunodeficiencies

“…A possibility is that these other staphylococci may share a mutualistic or commensal relationship with S. aureus by enabling greater resistance to antimicrobial peptides (Peschel et al 2001;SieprawskaLupa et al 2004;Lai et al 2007) or antibiotics (Wielders et al 2001;Berglund and Söderquist 2008) or by potentiating the S. aureus toxic response as with C. albicans (Peters and Noverr 2013). Alternately, these species could take advantage of inflamed skin conditions and co-colonize with S. aureus (Nilsson et al 1998;McCrea et al 2000;Cho et al 2001;Williams et al 2002).…”

Staphylococcus epidermidis device-related infections: pathogenesis and clinical management

“…Fbe, SdrG, SdrF and SdrH are all members of the recently identified protein family, the serine-aspartate (SD)-repeat-containing (Sdr) family of cellwall-anchored surface proteins (von Eiff et al 2002), which interact with extracellular matrix proteins. Embp, a fibronectin-binding protein (Williams et al 2002) and GehD, which binds to collagen (Bowden et al 2002), also specifically bind to extracellular matrix proteins and mediate adherence of S. epidermidis cells. In addition, the non-protein molecule techoic acid, found in the cell wall, interacts with immobilised fibronectin, mediating binding of S. epidermidis (Hussain et al 2001).…”