Specificity of the fucose-binding lectin of<i>Pseudomonas aeruginosa</i>

Garber, N.; Guempel, Ulrike M.; Gilboa‐Garber, Nechama; Royle, R.J.

doi:10.1111/j.1574-6968.1987.tb02619.x

Cited by 59 publications

(28 citation statements)

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“…The localization of LecB in the outer membrane of P. aeruginosa and its high affinity for L-fucose and its derivatives, like pNPF (Garber et al, 1987), suggested that this lectin might be bound to the outer membrane via fucose-containing structures. This assumption was tested by incubating an isolated outer-membrane fraction with 20 mM of either pNPF or D-galactose, which served as a negative control because the affinity of LecB for D-galactose is extremely low (Garber et al, 1987).…”

Section: Lecb Binds To Carbohydrate Ligands In the Outer Membranementioning

confidence: 99%

“…It consists of four 12?75 kDa subunits (Gilboa-Garber, 1972;Avichezer et al, 1992) and has been shown to cause cytotoxic effects on respiratory epithelial cells by decreasing their growth rate, thus contributing to respiratory epithelial injury during P. aeruginosa infection (Bajolet-Laudinat et al, 1994). In addition, it was demonstrated that LecA induces a permeability defect in the intestinal epithelium, resulting in increased absorption of exotoxin A, an important extracellular virulence factor of P. aeruginosa (Laughlin et al, 2000).LecB consists of four 11?73 kDa subunits, each exhibiting a high specificity for L-fucose and its derivatives (GilboaGarber et al, 2000;Garber et al, 1987) and also for Dmannose, with lower affinity. Its crystal structure was determined recently (Mitchell et al, 2002(Mitchell et al, , 2005Loris et al, 2003).…”

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Pseudomonas aeruginosa lectin LecB is located in the outer membrane and is involved in biofilm formation

et al. 2005

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Pseudomonas aeruginosa is an opportunistic pathogen which causes a variety of diseases, including respiratory tract infections in patients suffering from cystic fibrosis. Therapeutic treatment of P. aeruginosa infections is still very difficult because the bacteria exhibit high intrinsic resistance against a variety of different antibiotics and, in addition, form stable biofilms, e.g. in the human lung. Several virulence factors are produced by P. aeruginosa, among them the two lectins LecA and LecB, which exert different cytotoxic effects on respiratory epithelial cells and presumably facilitate bacterial adhesion to the airway mucosa. Here, the physiology has been studied of the lectin LecB, which binds specifically to L-fucose. A LecB-deficient P. aeruginosa mutant was shown to be impaired in biofilm formation when compared with the wild-type strain, suggesting an important role for LecB in this process. This result prompted an investigation of the subcellular localization of LecB by cell fractionation and subsequent immunoblotting. The results show that LecB is abundantly present in the bacterial outer-membrane fraction. It is further demonstrated that LecB could be released specifically by treatment of the outer-membrane fraction with p-nitrophenyl alpha-L-fucose, whereas treatment with D-galactose had no effect. In contrast, a LecB protein carrying the mutation D104A, which results in a defective sugar-binding site, was no longer detectable in the membrane fraction, suggesting that LecB binds to specific carbohydrate ligands located at the bacterial cell surface. Staining of biofilm cells using fluorescently labelled LecB confirmed the presence of these ligands.

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Section: Lecb Binds To Carbohydrate Ligands In the Outer Membranementioning

confidence: 99%

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Pseudomonas aeruginosa lectin LecB is located in the outer membrane and is involved in biofilm formation

et al. 2005

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“…solanacearum possesses a PA-IIL-like lectin (RS-IIL), but does not produce a PA-IL-like one (Gilboa-Garber et al, 2000;Sudakevitz et al, 2004). RS-IIL, which is shorter than PA-IIL by one amino acid (11?60 kDa), resembles it in its composition, Ca 2+ requirement for activity and a very high sugar affinity (Garber et al, 1987;Sudakevitz et al, 2004). However, it differs from PA-IIL in showing highest affinity to mannose, accompanied by a lower avidity for fucose and related sugars (Sudakevitz et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

“…aeruginosa produces PA-IIL (11?73 kDa) (Gilboa-Garber, 1982;Garber et al, 1987) in addition to a galactophilic lectin, PA-IL (12?76 kDa) (Gilboa-Garber, 1972). Both lectins are composed of four identical subunits (Mitchell et al, 2002;Ciocci et al, 2003;Imberty et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

Production and properties of the native Chromobacterium violaceum fucose-binding lectin (CV-IIL) compared to homologous lectins of Pseudomonas aeruginosa (PA-IIL) and Ralstonia solanacearum (RS-IIL)

et al. 2006

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Chromobacterium violaceum is a versatile, violet pigment (violacein)-producing b-proteobacterium, confined to tropical and subtropical regions, dwelling in soil and water, like Pseudomonas aeruginosa and Ralstonia solanacearum. These three bacteria are saprophytes that occasionally become aggressive opportunistic pathogens virulently attacking animals (the first two) and plants (the third). The recent availability of their genome sequences enabled identification in the C. violaceum genome of an ORF (locus no. 1744) that is similar to those of P. aeruginosa and R. solanacearum lectins, PA-IIL and RS-IIL, respectively. A recombinant protein, CV-IIL, encoded by that ORF exhibited fucose>mannose-specific lectin activity resembling PA-IIL. This paper describes production and properties of the native CV-IIL, which, like PA-IIL and RS-IIL, is probably also a quorum-sensing-driven secondary metabolite, appearing concomitantly with violacein. Its formation is repressed in the CV026 mutant of C. violaceum, which lacks endogenous N-acylhomoserine lactone. The upstream extragenic sequence of its ORF contains a 20 bp sequence (59-101-120) with partial similarities to the luxI-box and the related P. aeruginosa and R. solanacearum promoter boxes of quorum-sensing-controlled genes. The lectin level is augmented by addition of trehalose to the medium. The subunit size of CV-IIL (around 11?86 kDa) is similar to those of PA-IIL (11?73 kDa) and RS-IIL (11?60 kDa). Like PA-IIL, in the tetrameric form CV-IIL preferentially agglutinates a1-2 fucosylated H-positive human erythrocytes (regardless of their A, B or O type), as opposed to the O h Bombay type, but differs from it in having no interaction with rabbit erythrocytes and in displaying stronger affinity to L-galactose than to L-fucose. The greater similarity of CV-IIL to PA-IIL than to RS-IIL might be related to the selective adaptation of both C. violaceum and P. aeruginosa to animal tissues versus the preferential homing of R. solanacearum to plants. INTRODUCTIONChromobacterium violaceum is a versatile Gram-negative bproteobacterium (Dewhirst et al., 1989) that produces the violet non-diffusible antibiotic pigment violacein (Lichstein & Van de Sand, 1945;Antonio & Creczynski-Pasa, 2004) and additional antibiotics and enzymes affecting viruses, bacterial and eukaryotic cells (Duran & Menck, 2001;Melo et al., 2003;Chernin et al., 1998). This bacterium is a common inhabitant of soil and water confined to tropical and subtropical regions. Generally, it behaves as a saprophyte, but sporadically it becomes an aggressive opportunistic animal (including human) pathogen, causing serious infections with a high mortality in immunodeficient individuals (Pemberton et al., 1991; Shao et al., 2002;Alves de Brito et al., 2004). In that behaviour, C. violaceum resembles the green pigment (pyocyanin)-producing bacterium Pseudomonas aeruginosa, which is a worldwide inhabitant of similar ecosystems and has become an important common nosocomial pathogen, endangering hospitalized chronic patients. A...

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“…For example, in Pseudomonas aeruginosa, the matrix protein CdrA is both cell associated and secreted and likely promotes cell-matrix and matrix-matrix interactions by binding to the Psl polysaccharide (8), which itself is cell associated and secreted (9). Additionally, the lectins LecA, specific for D-galactose (10), and LecB, specific for L-fucose (11), are thought to promote cell-cell interactions (12) during P. aeruginosa biofilm formation.…”

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Identification of a Novel Matrix Protein That Promotes Biofilm Maturation in Vibrio fischeri

Ray¹,

Driks²,

Visick³

2014

J. Bacteriol.

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Bacteria form communities, termed biofilms, in which cells adhere to each other within a matrix, typically comprised of polysaccharides, proteins, and extracellular DNA. Biofilm formation by the marine bacterium Vibrio fischeri requires the Syp polysaccharide, but the involvement of matrix proteins is as yet unknown. Here we identified three genes, termed bmpA, -B, and -C (biofilm maturation protein), with overlapping functions in biofilm maturation. A triple bmpABC mutant, but not single or double mutants, was defective in producing wrinkled colonies, a form of biofilm. Surprisingly, the triple mutant was competent to form pellicles, another biofilm phenotype, but they generally lacked a three-dimensional architecture. Transmission electron microscopy revealed that the extracellular matrix of the bmp mutant contained electron-dense, thread-like structures that were also present in the wild type but lacking in syp mutant strains. We hypothesized that the bmp mutant produces the Syp polysaccharide but fails to produce/export a distinct matrix component. Indeed, a mixture of the bmp and syp mutants produced a wrinkled colony. Finally, BmpA could be detected in cell-free supernatants from disrupted pellicles. Thus, this work identifies a new matrix protein necessary for biofilm maturation by V. fischeri and, based on the conservation of bmp, potentially other microbes. Bacteria can be found as planktonic cells or within sessile communities called biofilms, which are composed of bacterial cells encased in an extracellular matrix (1-3). The matrix is generally self-produced and comprised of polysaccharides, proteins, extracellular DNA, and other bacterial products, such as outer membrane vesicles (OMVs). These matrix components promote adherence of the bacteria to each other and to a variety of surfaces, including host tissues and medical devices, such as catheters (reviewed in reference 4). Furthermore, due at least in part to the protective nature of the matrix, resident cells exhibit increased resistance to predation (5), desiccation (6), host defenses (4), and antibiotics (7). While much is known about the process of biofilm formation and its regulation from the study of a variety of bacteria, we still know little about the constituents of the matrix, in particular the matrix proteins, and their contributions to biofilm formation.Matrix proteins can be anchored to or associated with the cell surface or released into the extracellular space, where they appear to promote cell-matrix, cell-cell, matrix-matrix, and cell-surface interactions. For example, in Pseudomonas aeruginosa, the matrix protein CdrA is both cell associated and secreted and likely promotes cell-matrix and matrix-matrix interactions by binding to the Psl polysaccharide (8), which itself is cell associated and secreted (9). Additionally, the lectins LecA, specific for D-galactose (10), and LecB, specific for L-fucose (11), are thought to promote cell-cell interactions (12) during P. aeruginosa biofilm formation.Perhaps the most is known about the r...

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Specificity of the fucose-binding lectin ofPseudomonas aeruginosa

Cited by 59 publications

References 9 publications

Pseudomonas aeruginosa lectin LecB is located in the outer membrane and is involved in biofilm formation

Pseudomonas aeruginosa lectin LecB is located in the outer membrane and is involved in biofilm formation

Production and properties of the native Chromobacterium violaceum fucose-binding lectin (CV-IIL) compared to homologous lectins of Pseudomonas aeruginosa (PA-IIL) and Ralstonia solanacearum (RS-IIL)

Identification of a Novel Matrix Protein That Promotes Biofilm Maturation in Vibrio fischeri

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