The Fucose-binding Lectin from Ralstonia solanacearum

Plant pathogens, like animal ones, use protein-carbohydrate interactions in their strategy for host recognition, attachment, and invasion. The bacterium Ralstonia solanacearum, which is distributed worldwide and causes lethal wilt in many agricultural crops, was shown to produce a potent l-fucose-binding lectin, R. solanacearum lectin, a small protein of 90 amino acids with a tandem repeat in its amino acid sequence. In the present study, surface plasmon resonance experiments conducted on a series of oligosaccharides show a preference for binding to αFuc1-2Gal and αFuc1-6Gal epitopes. Titration microcalorimetry demonstrates the presence of two binding sites per monomer and an unusually high affinity of the lectin for αFuc1-2Gal-containing oligosaccharides (KD = 2.5 × 10-7 m for 2-fucosyllactose). R. solanacearum lectin has been crystallized with a methyl derivative of fucose and with the highest affinity ligand, 2-fucosyllactose. X-ray crystal structures, the one with α-methyl-fucoside being at ultrahigh resolution, reveal that each monomer consists of two small four-stranded anti-parallel β-sheets. Trimerization through a 3-fold or pseudo-3-fold axis generates a six-bladed β-propeller architecture, very similar to that previously described for the fungal lectin of Aleuria aurantia. This is the first report of a β-propeller formed by oligomerization and not by sequential domains. Each monomer presents two fucose binding sites, resulting in six symmetrically arranged sugar binding sites for the β-propeller. Crystals were also obtained for a mutated lectin complexed with a fragment of xyloglucan, a fucosylated polysaccharide from the primary cell wall of plants, which may be the biological target of the lectin.

[1]  J. Weissenbach,et al.  Genome sequence of the plant pathogen Ralstonia solanacearum , 2002, Nature.

[2]  M. Paoli,et al.  Protein folds propelled by diversity. , 2001, Progress in biophysics and molecular biology.

[3]  I. Wilson,et al.  Glycosylation of proteins in plants and invertebrates. , 2002, Current opinion in structural biology.

[4]  A. Imberty,et al.  A new Ralstonia solanacearum high‐affinity mannose‐binding lectin RS‐IIL structurally resembling the Pseudomonas aeruginosa fucose‐specific lectin PA‐IIL , 2004, Molecular microbiology.

[5]  J. Zou,et al.  Improved methods for building protein models in electron density maps and the location of errors in these models. , 1991, Acta crystallographica. Section A, Foundations of crystallography.

[6]  G. Chambat,et al.  Variation of xyloglucan substitution pattern affects the sorption on celluloses with different degrees of crystallinity , 2005 .

[7]  G. Bricogne,et al.  [27] Maximum-likelihood heavy-atom parameter refinement for multiple isomorphous replacement and multiwavelength anomalous diffraction methods. , 1997, Methods in enzymology.

[8]  R. Carlson,et al.  Oligosaccharins--oligosaccharides that regulate growth, development and defence responses in plants. , 1992, Glycobiology.

[9]  P. Albersheim,et al.  An unambiguous nomenclature for xyloglucan‐derived oligosaccharides , 1993 .

[10]  David T. Jones,et al.  β Propellers: structural rigidity and functional diversity , 1999 .

[11]  L. Wyns,et al.  Solving the phase problem for carbohydrate-binding proteins using selenium derivatives of their ligands: a case study involving the bacterial F17-G adhesin. , 2003, Acta crystallographica. Section D, Biological crystallography.

[12]  C. Boucher,et al.  A bacterial sensor of plant cell contact controls the transcriptional induction of Ralstonia solanacearum pathogenicity genes , 2000, The EMBO journal.

[13]  P. Kraulis.  A program to produce both detailed and schematic plots of protein structures , 1991 .

[14]  M. Schell.  Control of Virulence and Pathogenicity Genes of Ralstonia Solanacearum by an Elaborate Sensory Network. , 2000, Annual review of phytopathology.

[15]  K. Miki,et al.  Crystal structure of fucose-specific lectin from Aleuria aurantia binding ligands at three of its five sugar recognition sites. , 2003, Biochemistry.

[16]  A. Hayward.  Biology and epidemiology of bacterial wilt caused by pseudomonas solanacearum. , 1991, Annual review of phytopathology.

[17]  Jaroslav Koča,et al.  Computer simulation of histo-blood group oligosaccharides: energy maps of all constituting disaccharides and potential energy surfaces of 14 ABH and Lewis carbohydrate antigens , 1995, Glycoconjugate Journal.

[18]  K. Ruel,et al.  Structural features and biological activity of xyloglucans from suspension-cultured plant cells. , 1992, Biochimie.

[19]  A. Imberty,et al.  Production, properties and specificity of a new bacterial L-fucose- and D-arabinose-binding lectin of the plant aggressive pathogen Ralstonia solanacearum, and its comparison to related plant and microbial lectins. , 2002, Journal of biochemistry.

[20]  J. Simpson,et al.  Oligosaccharides related to xyloglucan: synthesis and X-ray crystal structure of methyl α-l-fucopyranosyl-(1→2)-β-d-galactopyranosyl-(1→2)-α-d-xylopyranoside and the synthesis of methyl α-l-fucopyranosyl-(1→2)-β-d-galactopyranosyl-(1→2)-β-d-xylopyranoside , 2000 .

[21]  C. Verlinde,et al.  Structural biology and structure-based inhibitor design of cholera toxin and heat-labile enterotoxin. , 2004, International journal of medical microbiology : IJMM.

[22]  Anastassis Perrakis,et al.  Automated protein model building combined with iterative structure refinement , 1999, Nature Structural Biology.

[23]  R. Huber,et al.  Tachylectin‐2: crystal structure of a specific GlcNAc/GalNAc‐binding lectin involved in the innate immunity host defense of the Japanese horseshoe crab Tachypleus tridentatus , 1999, The EMBO journal.

[24]  Collaborative Computational,et al.  The CCP4 suite: programs for protein crystallography. , 1994, Acta crystallographica. Section D, Biological crystallography.

[25]  J. Thornton,et al.  PROCHECK: a program to check the stereochemical quality of protein structures , 1993 .

[26]  J. Vincken,et al.  Potato xyloglucan is built from XXGG-type subunits. , 1996, Carbohydrate research.

[27]  Anne Imberty,et al.  Crystal Structure of Fungal Lectin , 2003, Journal of Biological Chemistry.

[28]  P. Albersheim,et al.  Structure and function of the primary cell walls of plants. , 1984, Annual review of biochemistry.

[29]  G. Murshudov,et al.  Refinement of macromolecular structures by the maximum-likelihood method. , 1997, Acta crystallographica. Section D, Biological crystallography.

[30]  N. Gilboa-Garber.  Pseudomonas aeruginosa lectins. , 1982, Methods in enzymology.

[31]  Y. Hata,et al.  Molecular Cloning and Overexpression of fleA Gene Encoding a Fucose-specific Lectin of Aspergillus oryzae , 2002, Bioscience, biotechnology, and biochemistry.

[32]  K. Acharya,et al.  Botulinum and tetanus neurotoxins: structure, function and therapeutic utility. , 2002, Trends in biochemical sciences.

[33]  J. Simpson,et al.  Oligosaccharides related to xyloglucan: Synthesis and X-ray crystal structure of methyl 2-O-(α-l-fucopyranosyl)-β-d-galactopyranoside , 1996 .

[34]  Martin Knoblauch,et al.  MOLCAD — Computer Aided Visualization and Manipulation of Models in Molecular Science , 1992 .

[35]  G Waksman,et al.  Chaperone-assisted pilus assembly and bacterial attachment. , 2000, Current opinion in structural biology.

[36]  P. Albersheim,et al.  The structures of arabinoxyloglucans produced by solanaceous plants. , 1996, Carbohydrate research.

[37]  J. Gordon,et al.  Glycans as legislators of host-microbial interactions: spanning the spectrum from symbiosis to pathogenicity. , 2001, Glycobiology.

[38]  T. Dam,et al.  Thermodynamic studies of lectin-carbohydrate interactions by isothermal titration calorimetry. , 2002, Chemical reviews.

[39]  S. Teneberg,et al.  Interaction of viruses, bacteria and bacterial toxins with host cell surface glycolipids. Aspects on receptor identification and dissection of binding epitopes. , 1988, Advances in experimental medicine and biology.

[40]  A. Otter,et al.  Crystal state and solution conformation of the B blood group trisaccharide α-l-Fuc p -(1→2)-[α-d-Gal p ]-(1→3)]-β-d-Gal p -OCH 3 , 1999 .