The SasG surface protein of Staphylococcus aureus has been shown to promote the formation biofilm. comprises an N-terminal A domain and repeated B domains. Here we demonstrate that is involved in accumulation phase biofilm, a process requires physiological concentration Zn(2+). domains, but not domain, are required. Purified recombinant can form dimers vitro Zn(2+)-dependent fashion. Furthermore, bind cells have domains anchored their block biofilm formation. full-length exposed on cell...

Entry of Staphylococcus aureus into the bloodstream can lead to metastatic abscess formation and infective endocarditis. Crucial development both these conditions is interaction S. with endothelial cells. In vivo in vitro studies have shown that staphylococcal invasin FnBPA triggers bacterial invasion cells via a process involves fibronectin (Fn) bridging α5β1 integrins. The Fn-binding region usually contains 11 non-identical repeats (FnBRs) differing affinities for Fn, which facilitate...

Staphylococcus aureus can adhere to and invade endothelial cells by binding the human protein fibronectin (Fn). FnBPA FnBPB, cell wall-attached proteins from S. aureus, have multiple, intrinsically disordered, high-affinity repeats (FnBRs) for Fn. Here, 30 years after first report of aureus/Fn interactions, we present four crystal structures that together comprise two complete FnBRs, each in complex with N-terminal modules Each approximately 40-residue FnBR forms antiparallel strands along...

Enzymatic cleavage of the glycosidic bond yields products in which anomeric configuration is either retained or inverted. Each mechanism reflects dispositions enzyme functional groups; a facet essentially conserved 113 glycoside hydrolase (GH) families. We show that family GH97 has diverged significantly, as it contains both inverting and retaining α-glycosidases. This evolution active center; glutamate acts general base members, exemplified by Bacteroides thetaiotaomicron α-glucosidase...

Staphylococcus aureus and epidermidis form communities (called biofilms) on inserted medical devices, leading to infections that affect many millions of patients worldwide cause substantial morbidity mortality. As biofilms are resistant antibiotics, device removal is often required resolve the infection. Thus, there a need for new therapeutic strategies molecular data might assist their development. Surface proteins S. surface protein G (SasG) accumulation-associated (S. epidermidis) promote...

The explosive 2,4,6-trinitrotoluene (TNT) is a major worldwide military pollutant. presence of this toxic and highly persistent pollutant, particularly at sites former manufacturing facilities, presents various health environmental concerns. Due to the chemically resistant structure TNT, it has proven be recalcitrant biodegradation in environment. Here, we demonstrate importance two glutathione transferases (GSTs), GST-U24 GST-U25, from Arabidopsis (Arabidopsis thaliana) that are...

Binding of the fibronectin-binding protein FnBPA from Staphylococcus aureus to human fibronectin has previously been implicated in development infective endocarditis, specifically processes platelet activation and invasion endothelium. We recently proposed a model for binding which bacterial contains 11 potential sites (FnBPA-1 FnBPA-11), each composed motifs that bind consecutive type 1 modules N-terminal domain fibronectin. Here we show six with dissociation constants nanomolar range;...

Staphylococcus aureus is a commensal organism and frequent cause of skin soft tissue infections, which can progress to serious invasive disease. This bacterium uses its fibronectin binding proteins (FnBPs) invade host cells it has been hypothesised that this provides protected niche from antimicrobial defences, allows access deeper tissues reservoir for persistent or recurring infections. FnBPs contain multiple tandem fibronectin-binding repeats (FnBRs) bind with varying affinity but unclear...

Abstract Bacteria exploit surface proteins to adhere other bacteria, surfaces and host cells. Such need project away from the bacterial resist significant mechanical forces. SasG is a protein that forms extended fibrils on of Staphylococcus aureus promotes adherence biofilm formation. Here we show although monomeric lacking covalent cross-links, maintains highly conformation in solution. This extension mediated through obligate folding cooperativity intrinsically disordered E domains couple...

Adherence of microbes to host tissues is a hallmark infectious disease and often mediated by class adhesins termed MSCRAMMs (Microbial Surface Components Recognizing Adhesive Matrix Molecules). Numerous pathogens express that specifically bind the heterodimeric human glycoprotein fibronectin (Fn). In addition roles in adhesion, Fn-binding exploit physiological Fn functions. For example, several can invade cells mechanism whereby MSCRAMM-bound bridges interaction with α5β1 integrin. Here, we...

Collision cross-section (CCS) measurements obtained from ion mobility spectrometry-mass spectrometry (IMS-MS) analyses often provide useful information concerning a protein's size and shape can be complemented by modeling procedures. However, there have been some concerns about the extent to which certain proteins maintain native-like conformation during gas-phase analysis, especially with dynamic or extended regions. Here we measured CCSs of range biomolecules including non-globular RNAs...

BBK32 is a fibronectin-binding protein from the Lyme disease-causing spirochete Borrelia burgdorferi. In this study, we show that shares sequence similarity with fibronectin module-binding motifs previously identified in proteins Streptococcus pyogenes and Staphylococcus aureus. Nuclear magnetic resonance spectroscopy isothermal titration calorimetry are used to confirm binding sites of peptides within N-terminal domain measure affinities interactions. Comparison chemical shift perturbations...

BBK32 is a fibronectin-binding lipoprotein on Borrelia burgdorferi, the causative agent of Lyme disease. Analysis using secondary structure prediction programs suggested that composed two domains, an N-terminal segment lacking well defined and C-terminal largely alpha-helices. purified recombinant forms domains by circular dichroism spectroscopy, gel permeation chromatography, intrinsic viscosity determination were consistent with N-terminal-extended, unstructured globular domain in BBK32....

Staphylococcus aureus and epidermidis cause dangerous difficult to treat medical device‐related infections through their ability form biofilms. Extracellular poly‐ N ‐acetylglucosamine (PNAG) facilitates biofilm formation is a vaccination target, yet details of its biosynthesis by the icaADBC gene products limited. IcaC proposed transporter for PNAG export, however comparison Ica proteins homologous exo‐polysaccharide synthases suggests that common IcaAD protein components both synthesise...

Fibronectin (Fn) binding by the Streptococcus pyogenes protein SfbI has been shown to trigger integrin-dependent internalization of this pathogen human epithelial and endothelial cells. Here, using nuclear magnetic resonance spectroscopy isothermal titration calorimetry in a dissection approach, basis for specificity high affinity interaction between N-terminal domain Fn is revealed. Each five type 1 modules directly involved recognized short consecutive motifs within repeat region SfbI....

The effect of osmotic cell swelling on the permeability HeLa cells to a range structurally unrelated solutes including taurine, sorbitol, thymidine, choline, and K+ (96Rb+) was investigated. For each solute tested, reduction in osmolality medium from 300 200 mosmol/kgH2O caused significant increase unidirectional influx rate. In case, osmotically activated transport component nonsaturable up external substrate concentrations 50 mM. Inhibitors swelling-activated anion channel [quinine,...

The acquisition of host-derived sialic acid is an important virulence factor for some bacterial pathogens, but in vivo this sugar sequestered sialoconjugates as the alpha-anomer. In solution, however, present mainly beta-anomer, formed by a slow spontaneous mutarotation. We studied Escherichia coli protein YjhT member family uncharacterized proteins many acid-utilizing pathogens. This able to accelerate equilibration alpha- and beta-anomers N-acetylneuraminic acid, thus describing novel...

Many human proteins contain intrinsically disordered regions, and disorder in these can be fundamental to their function-for example, facilitating transient but specific binding, promoting allostery, or allowing efficient posttranslational modification. SasG, a multidomain protein implicated host colonization biofilm formation Staphylococcus aureus, provides another example of how play an important role. Approximately one-half the domains extracellular repetitive region SasG are unfolded...