Identification of glycoprotein receptors within the human salivary proteome for the lectin‐like BabA and SabA adhesins of <b><i>Helicobacter pylori</i></b> by fluorescence‐based 2‐D bacterial overlay

Walz, Anke; Odenbreit, Stefan; Stühler, Kai; Wattenberg, Andreas; Meyer, Helmut E.; Mahdavi, Jafar; Borén, Thomas; Rühl, Stefan

doi:10.1002/pmic.200700808

Cited by 44 publications

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“…Yet, our knowledge on whether and how medical pathogens are affected by the salivary proteome during their passage through or residence within the mouth is limited (20)(21)(22)(23)(24). In this study, we have demonstrated that S. aureus rapidly and selectively binds a limited number of salivary proteins to its surface following exposure to saliva.…”

Section: Discussionmentioning

confidence: 99%

“…Studies have shown the occurrence of S. aureus in oral biofilm and saliva of healthy individuals (18), but its frequency was found higher in elderly and institutionalized individuals, including hospitalized and nursing home patients (9,19). Yet, in spite of the well-described associations between salivary dysfunction, biofilm formation, and bacterial colonization, only a few studies have investigated the adhesive interactions of salivary components with medical pathogens, in particular S. aureus (20)(21)(22)(23)(24). Here, as a first step toward understanding the mechanism by which pathogens can colonize the oral cavity of vulnerable patients, S. aureus was chosen as a model organism to identify specific salivary components that bind to the bacterium and to elucidate the role of biofilm formation for the bacterium's ability to bind salivary proteins.…”

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confidence: 99%

“…Parotid saliva and submandibular-sublingual saliva were collected following stimulation of salivary flow by application of 2% citric acid to the dorsum of the tongue. Parotid saliva was collected from the orifice of Stensons's duct using a modified Carlsen-Crittenden device, and submandibular-sublingual saliva was collected from the floor of the mouth with disposable plastic transfer pipettes (VWR International, Radnor, PA) as previously described (21). Protein concentrations of clarified WS were determined using the Pierce bicinchoninic acid (BCA) protein assay kit (Thermo Scientific, Rockland, IL), with bovine serum albumin (BSA; fraction V; Sigma, St. Louis, MO) as the standard.…”

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Host Defense Proteins Derived from Human Saliva Bind to Staphylococcus aureus

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bProteins in human saliva are thought to modulate bacterial colonization of the oral cavity. Yet, information is sparse on how salivary proteins interact with systemic pathogens that transiently or permanently colonize the oral environment. Staphylococcus aureus is a pathogen that frequently colonizes the oral cavity and can cause respiratory disease in hospitalized patients at risk. Here, we investigated salivary protein binding to this organism upon exposure to saliva as a first step toward understanding the mechanism by which the organism can colonize the oral cavity of vulnerable patients. By using fluorescently labeled saliva and proteomic techniques, we demonstrated selective binding of major salivary components by S. aureus to include DMBT1 gp-340 , mucin-7, secretory component, immunoglobulin A, immunoglobulin G, S100-A9, and lysozyme C. Biofilm-grown S. aureus strains bound fewer salivary components than in the planctonic state, particularly less salivary immunoglobulins. A corresponding adhesive component on the S. aureus surface responsible for binding salivary immunoglobulins was identified as staphylococcal protein A (SpA). However, SpA did not mediate binding of nonimmunoglobulin components, including mucin-7, indicating the involvement of additional bacterial surface adhesive components. These findings demonstrate that a limited number of salivary proteins, many of which are associated with various aspects of host defense, selectively bind to S. aureus and lead us to propose a possible role of saliva in colonization of the human mouth by this pathogen. Saliva plays a key role in host defense against invading pathogens (1-4). Among the more than 2,000 proteins and peptides found in saliva (5), many exhibit direct antimicrobial activity (6). Others can bind to bacteria to facilitate either their colonization on oral surfaces or their clearance from the oral cavity through agglutination (7,8). It has been suggested that systemic pathogens can be killed, inactivated, or agglutinated by salivary components and, thus, become cleared from the oral cavity through swallowing, thereby preventing them from colonizing the oral cavity of healthy individuals (2, 9). Thus, binding of salivary proteins to pathogens is thought to play an important role in preventing systemic infections. In hospitalized patients, the protective and antimicrobial functions of saliva, which play a crucial role in host defense against invading pathogens (1-3), are frequently impaired by reduction of salivary flow or lack of salivary secretion (9-11). Under such conditions of dry mouth and poor oral hygiene, the normal commensal oral microflora shifts to a community that harbors a higher number of pathogens (12, 13).Among the various systemic pathogens in the oral cavity, attention has been given to Staphylococcus aureus (14, 15), since both endocarditis and pneumonia have been related to oral colonization by this organism (16,17). Studies have shown the occurrence of S. aureus in oral biofilm and saliva of healthy individuals (18), bu...

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Host Defense Proteins Derived from Human Saliva Bind to Staphylococcus aureus

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“…Using this strategy, it was demonstrated, as an example, that binding to the salivary mucin MUC5B is mainly mediated by the BabA adhesin [20]. Purified glycoproteins and neoglycoproteins immobilized on arrays have also been used as a tool for discovery and characterization of new bacterial lectin-carbohydrate interactions [21,22].…”

Section: Discovering Other H Pylori Sweet Interactions In Gastric Mumentioning

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Sweet receptors mediate the adhesion of the gastric pathogenHelicobacter pylori: glycoproteomic strategies

Magalhães

Ismail

Reis³

2010

Expert Review of Proteomics

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“…For this reason, we have adapted here the bacterial overlay method (13,14) and combined it with a method for immobilizing bacteria (15) to allow efficient screening of multispecies biofilms for coadhesion partners with the option to isolate and identify them subsequently. The resulting coadhesion assay was first validated using strains of oral actinomyces and streptococci that are known to coaggregate in vitro (8,9) and engage in coadhesion in vivo (12).…”

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Probing of Microbial Biofilm Communities for Coadhesion Partners

Rühl

Eidt

Melzl

et al. 2014

Appl Environ Microbiol

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dInvestigations of interbacterial adhesion in dental plaque development are currently limited by the lack of a convenient assay to screen the multitude of species present in oral biofilms. To overcome this limitation, we developed a solid-phase fluorescencebased screening method to detect and identify coadhesive partner organisms in mixed-species biofilms. The applicability of this method was demonstrated using coaggregating strains of type 2 fimbrial adhesin-bearing actinomyces and receptor polysaccharide (RPS)-bearing streptococci. Specific adhesin/receptor-mediated coadhesion was detected by overlaying bacterial strains immobilized to a nitrocellulose membrane with a suspended, fluorescein-labeled bacterial partner strain. Coadhesion was comparable regardless of which cell type was labeled and which was immobilized. Formaldehyde treatment of bacteria, either in suspension or immobilized on nitrocellulose, abolished actinomyces type 2 fimbrial adhesin but not streptococcal RPS function, thereby providing a simple method for assigning complementary adhesins and glycan receptors to members of a coadhering pair. The method's broader applicability was shown by overlaying colony lifts of dental plaque biofilm cultures with fluoresceinlabeled strains of type 2 fimbriated Actinomyces naeslundii or RPS-bearing Streptococcus oralis. Prominent coadhesion partners included not only streptococci and actinomyces, as expected, but also other bacteria not identified in previous coaggregation studies, such as adhesin-or receptor-bearing strains of Neisseria pharyngitis, Rothia dentocariosa, and Kingella oralis. The ability to comprehensively screen complex microbial communities for coadhesion partners of specific microorganisms opens a new approach in studies of dental plaque and other mixed-species biofilms.

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Identification of glycoprotein receptors within the human salivary proteome for the lectin‐like BabA and SabA adhesins of Helicobacter pylori by fluorescence‐based 2‐D bacterial overlay

Cited by 44 publications

References 44 publications

Host Defense Proteins Derived from Human Saliva Bind to Staphylococcus aureus

Host Defense Proteins Derived from Human Saliva Bind to Staphylococcus aureus

Sweet receptors mediate the adhesion of the gastric pathogenHelicobacter pylori: glycoproteomic strategies

Probing of Microbial Biofilm Communities for Coadhesion Partners

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