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)

Abstract

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...

Preventing Pseudomonas aeruginosa and Chromobacterium violaceum infections by anti-adhesion-active components of edible seeds

“…P. aeruginosa produces a galactophilic lectin PA-IL (LecA) and a fucophilic (+ mannophilic and arabinophilic) lectin PA-IIL (LecB) [3]. C. violaceum also possesses a fucophilic lectin CV-IIL homologous to PA-IIL in structure and major specificity [4]. These three lectins bind to most human cells due to their affinities to their most common antigens [5,6]: PA-IL preferentially binds to the terminal Galα-bearing human blood group epitopes found in P-system, I, and B antigens [5].…”

Analyses of diverse mammals’ milk and lactoferrin glycans using five pathogenic bacterial lectins

“…CV-IIL was isolated from C. violaceum (Bergonzini) ATCC 12472, and RSL and RS-IIL -from R. solanacearum ATCC 11696 (purchased from the American Type Culture Collection, ATCC, Manassas, VA), as earlier described (GilboaGarber, 1982;Sudakevitz et al, 2002;Sudakevitz et al, 2004;Zinger-Yosovich et al, 2006). Con A and UEA-I were purchased from Sigma-Aldrich (St. Louis, MO).…”

“…They reported that blood-group H-type 1 precursor was expressed in all species investigated [camel, cow, goat, horse (mare), human, pig, rabbit and sheep (ewe)], while only pig, dromedary camel, and rabbit milk proteins carried the H-type 2 epitope, which is important for the inhibition of the adhesion of Helicobacter pylori, Escherichia coli heat-stable toxin, and Campylobacter jejuni to human cells. We have recently used three bacterial lectins (PA-IL, PA-IIL and CV-IIL) of the animal pathogens P. aeruginosa (Gilboa-Garber, 1982) and Chromobacterium violaceum (C. violaceum) (Zinger-Yosovich, Sudakevitz, Imberty, Garber, & Gilboa-Garber, 2006) for the study of diverse mammal milks (Zinger-Yosovich, Iluz, Sudakevitz, & Gilboa-Garber, 2010), showing their different lectin-blocking capacities.…”

“…The five pathogenic bacterial lectins used in this study vary in their carbohydrate specificities: PA-IL (LecA) is galactophilic and PA-IIL (LecB) is fucophilic (Gilboa-Garber, 1982); CV-IIL, which is also fucophilic, is a structural homologue of PA-IIL, exhibiting more selective H blood group specificity (Zinger-Yosovich et al, 2006); RSL, which is also fucophilic, exhibits Fuca1-2/3/4/6(core)-binding affinities (Kostlanova et al, 2005;Sudakevitz et al, 2002;Wu et al, 2009), and RS-IIL, which is another structural homologue of PA-IIL and CV-IIL, is mannophilic (Sudakevitz et al, 2004). These lectins are considered as virulence factors (VIFs) contributing to the bacterial adhesion to the target cells, to biofilm formation and to augmentation of the notorious effects of other VIFs (toxins and hydrolytic enzymes) which are co-produced with them (GilboaGarber et al, 1997).…”

Crystallization and preliminary X-ray diffraction analysis of an anti-H(O) lectin fromLotus tetragonolobusseeds

“…Fucose-binding lectins are widespread among microorganisms, animals and plants, including Pseudomonas aeruginosa lectin (PA-IIL), Anguilla anguilla agglutinin (AAA), Morone saxatilis agglutinin (MsaFBP32), Dicentrarchus labrax agglutinin, Chromobacterium violaceum lectin (CV-IIL), Ralstonia solanacearum lectin (RS-IIL), Ulex europaeus agglutinin (UEA-I) and Lotus tetragonolobus agglutinin (LTA) (Bianchet et al, 2002;Mitchell et al, 2005;Vandonselaar & Delbaere, 1994;Konami et al, 1990;Zinger-Yosovich et al, 2006;Odom & Vasta, 2006;Cammarata et al, 2001).…”