Adherence of actinomyces pyogenes to HeLa cells mediated by hydrophobic surface proteins

Ding, H.; Lämmler, Christoph; Seleim, R. S.

doi:10.1016/s0934-8840(11)80362-2

Cited by 7 publications

(8 citation statements)

References 27 publications

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“…In order to determine whether this was the case for A. pyogenes, experiments assessing the adhesion of NANH-1, the neuraminidase mutant, were conducted. As previously reported, A. pyogenes can adhere to HeLa cells (15), and we demonstrated adherence to other epithelial cell lines, although A. pyogenes adhered best to HeLa cells. NANH-1 displayed no defect in adherence to HeLa cells compared with the wild-type, BBR1.…”

Section: Discussionsupporting

confidence: 88%

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Cloning, Expression, and Characterization of a Neuraminidase Gene fromArcanobacterium pyogenes

2001

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Arcanobacterium pyogenes is an opportunistic pathogen, associated with suppurative infections in domestic animals. In addition to pyolysin, a pore-forming, cholesterol-binding toxin, A. pyogenes expresses a number of putative virulence factors, including several proteases and neuraminidase activity. A 3,009-bp gene, nanH, was cloned and sequenced and conferred neuraminidase activity on an Escherichia coli host strain. The predicted 107-kDa NanH protein displayed similarity to a number of bacterial neuraminidases and contained the RIP/RLP motif and five copies of the Asp box motif found in all bacterial neuraminidases. Recombinant His-tagged NanH was found to have pH and temperature optima of 5.5 to 6.0 and 55°C, respectively. Insertional deletion of the nanH gene resulted in the reduction, but not absence, of neuraminidase activity, indicating the presence of a second neuraminidase gene in A. pyogenes. NanH was localized to the A. pyogenes cell wall. A. pyogenes adhered to HeLa, CHO, and MDBK cells in a washing-resistant manner. However, the nanH mutant was not defective for adherence to epithelial cells. The role of NanH in host epithelial cell adherence may be masked by the presence of a second neuraminidase in A. pyogenes.Arcanobacterium pyogenes is a common inhabitant of the upper respiratory, urogenital (12, 54), and gastrointestinal tracts (35; B. H. Jost, K. W. Post, and S. J. Billington, unpublished data) of many domestic animal species. However, a physical or microbial insult to the host can lead to a variety of suppurative A. pyogenes infections, such as mastitis in dairy cows (27) and goats (2), liver abscesses in feedlot cattle (31, 34), and pneumonia in pigs (26) and various species of wildlife (17,42,57). A. pyogenes can also infect avian species (10) and humans (5,16,19), although infections in humans are rare.A. pyogenes elaborates a number of extracellular proteins, including the hemolytic exotoxin pyolysin (PLO) (8), several proteases (48, 51), a DNase (30), and at least one neuraminidase (47). While all these proteins are putative virulence factors, only for PLO is there definitive evidence of involvement in the pathogenesis of infections by A. pyogenes (29).Recently, there has been interest in the neuraminidases of bacterial pathogens and the potential role they play in pathogenesis. Neuraminidase (N-acetylneuraminyl hydrolase; EC 3.2.1.18) removes sialic acid from glycolipids, glycoproteins, and poly-and oligosaccharides. Bacterial neuraminidases have only 20 to 30% amino acid sequence identity, but they contain two conserved motifs, the RIP/ RLP motif (Arg-Ile/Leu-Pro) and the Asp box motif (Ser-X-Asp-X-Gly-X-Thr-Trp), which occurs four or five times in the enzyme (14,21,43).Neuraminidases are virulence factors, especially in bacteria that inhabit mucosal surfaces (20, 22, 52, 55), and they may play several roles in virulence. This enzyme can make sialic acid available as a carbon source to promote growth in a nutrient-limited environment (11, 23). The action of neuraminidase can decrease m...

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Section: Discussionsupporting

confidence: 88%

“…It was previously demonstrated that A. pyogenes adhered to HeLa cells (15). We also tested the ability of BBR1 to adhere to other epithelial cell lines.…”

Section: A Pyogenes Strains Express Neuraminidase Activitymentioning

confidence: 97%

Cloning, Expression, and Characterization of a Neuraminidase Gene fromArcanobacterium pyogenes

2001

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“…The presence of particular O antigens on the surface of P. aeruginosa is known to affect the overall charge and physicochemistry of the bacterial cell; strains lacking the B-band O antigen have demonstrated greater ability to adhere to abiotic hydrophobic surfaces (2). It is also known that other bacterial species exhibiting a hydrophobic surface demonstrate enhanced adhesion to epithelial cells (4,28). Therefore, it is likely that the loss of the B-band O antigen could facilitate acute infection via a number of cooperative mechanisms, including increased TTSS expression and increased host cell adherence.…”

Section: Discussionmentioning

confidence: 99%

Presence or Absence of Lipopolysaccharide O Antigens Affects Type III Secretion by Pseudomonas aeruginosa

Augustin

Song²,

Baek³

et al. 2007

J Bacteriol

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Pseudomonas aeruginosa is one of the major causative agents of mortality and morbidity in hospitalized patients due to a multiplicity of virulence factors associated with both chronic and acute infections. Acute P. aeruginosa infection is primarily mediated by planktonic bacteria expressing the type III secretion system (TTSS), a surface-attached needle-like complex that injects cytotoxins directly into eukaryotic cells, causing cellular damage. Lipopolysaccharide (LPS) is the principal surface-associated virulence factor of P. aeruginosa. This molecule is known to undergo structural modification (primarily alterations in the

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“…It is pertinent to note that Rhodococci have a unique cell wall which is characterized by thick layers of micolic acids to which proteins are attached mainly by hydrophobic interactions [22]. The susceptibility of the protein to extraction by mutanolysin, an enzyme which is known to release cell-wall proteins from Gram positive bacteria, indicates that TiBP is indeed extracellular.…”

Section: Discussionmentioning

confidence: 99%

“…GIN1 bacterium, intact bacterial cells were treated with mutanolysin [19], a muralytic enzyme that splits the beta 1-4 N-acetylmuramy1-N-acetylgucosamine linkage in the peptide-glycan layer, leading to the release of cell wall proteins of Gram positive bacteria including Rhodococci [21]. Ding et al [22] also reported the use of this enzyme to release cell-surface hydrophobic proteins from Actinomyces pyogenes. After removal of the cell debris by centrifugation, and separation of the titanium binding proteins with TiO 2 (anatase) Sachtopore column (eluted with 8 M urea containing 1 % SDS), three protein bands were detected by PAGE-SDS.…”

Section: Evidence For the Bacterial Cell-wall Location Of Tibpmentioning

confidence: 99%

A TiO₂‐Binding Protein isolated from Rhodococcus Strain GIN‐1 (NCIMB 40340) – Purification, Properties and Potential Applications

Gertler

Brudo

Kenig

et al. 2003

Materialwissenschaft Werkst

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A Rhodocuccus strain (Rh. GIN1, NCIMB 40340), which is capable of adsorption to titanium dioxide (TiO 2 ) and TiO 2 -containing coal fly ash particles, has been isolated previously in our Lab. Selectivity experiments showed that the bacterium is capable of adsorption to other metal oxides as well (e.g. magnetite and Al 2 O 3 ) but at lower affinities. The bacterium binds tightly both to rutile and anatase TiO 2 . In electronmicrograms the formation of "bridgelike" structures between the bacterium and the oxide is observed. A specific protein fraction, located on the cell wall of the bacterium was isolated from the bacterium. This protein was found to adhere strongly to TiO 2 particles at high salt concentrations, similarly to the binding to TiO 2 of the intact bacteria. TiO 2 (rutile) was found to bind the protein faster, stronger and at a higher capacity than the anatase isoform. The 55 kDa Ti-Binding Protein (TiBP) was isolated from the bacteria after homogenization by French Press. It was purified by affinity chromatography on TiO 2 particles, hydrophobic chromatography on a Fractogel-propyl column and gel filtration on a Superdex G-200 column. The same protein was isolated from the bacteria by treatment with mutanolysin, an enzyme which is commonly used to retrieve cell-wall proteins from Gram-positive bacteria, demonstrating the outer cell location of the protein in Rh. GIN1. TiBP exhibits metal oxide binding selectivity similar to that of the intact bacterium, namely TiO 2 >ZnO>Al 2 O 3 >Fe 2 O 3 (magnetite). Hydrophobic forces seem to dominate the interactions of the protein with TiO 2 as its binding capability is greatly enhanced in the presence of high concentrations of NaCl and its desorption requires high concentrations of urea and SDS. These features differentiate TiBP from other proteins known to adsorb TiO 2 (such as hemoglobin, cytochrome c and bovine serum albumin), mainly by weak, charge-based interactions.

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Adherence of actinomyces pyogenes to HeLa cells mediated by hydrophobic surface proteins

Cited by 7 publications

References 27 publications

Cloning, Expression, and Characterization of a Neuraminidase Gene fromArcanobacterium pyogenes

Cloning, Expression, and Characterization of a Neuraminidase Gene fromArcanobacterium pyogenes

Presence or Absence of Lipopolysaccharide O Antigens Affects Type III Secretion by Pseudomonas aeruginosa

A TiO₂‐Binding Protein isolated from Rhodococcus Strain GIN‐1 (NCIMB 40340) – Purification, Properties and Potential Applications

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Adherence of actinomyces pyogenes to HeLa cells mediated by hydrophobic surface proteins

Cited by 7 publications

References 27 publications

Cloning, Expression, and Characterization of a Neuraminidase Gene fromArcanobacterium pyogenes

Cloning, Expression, and Characterization of a Neuraminidase Gene fromArcanobacterium pyogenes

Presence or Absence of Lipopolysaccharide O Antigens Affects Type III Secretion by Pseudomonas aeruginosa

A TiO2‐Binding Protein isolated from Rhodococcus Strain GIN‐1 (NCIMB 40340) – Purification, Properties and Potential Applications

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Product

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About

A TiO₂‐Binding Protein isolated from Rhodococcus Strain GIN‐1 (NCIMB 40340) – Purification, Properties and Potential Applications