Galactose Recognition by the Apicomplexan Parasite Toxoplasma gondii*

Background: TgMIC4 is an important microneme effector protein from Toxoplasma gondii. Results: The structure of TgMIC4 together with carbohydrate microarray analyses reveal a broad specificity for galactose-terminating sequences. Conclusion: Lectin activity within the fifth apple domain of TgMIC4 is reminiscent of the mammalian galectin family. Significance: TgMIC4 may contribute to parasite dissemination within the host or down-regulation of the immune response. Toxosplasma gondii is the model parasite of the phylum Apicomplexa, which contains numerous obligate intracellular parasites of medical and veterinary importance, including Eimeria, Sarcocystis, Cryptosporidium, Cyclospora, and Plasmodium species. Members of this phylum actively enter host cells by a multistep process with the help of microneme protein (MIC) complexes that play important roles in motility, host cell attachment, moving junction formation, and invasion. T. gondii (Tg)MIC1-4-6 complex is the most extensively investigated microneme complex, which contributes to host cell recognition and attachment via the action of TgMIC1, a sialic acid-binding adhesin. Here, we report the structure of TgMIC4 and reveal its carbohydrate-binding specificity to a variety of galactose-containing carbohydrate ligands. The lectin is composed of six apple domains in which the fifth domain displays a potent galactose-binding activity, and which is cleaved from the complex during parasite invasion. We propose that galactose recognition by TgMIC4 may compromise host protection from galectin-mediated activation of the host immune system.

[1]  W. Chai,et al.  The human epithelial carcinoma antigen recognized by monoclonal antibody AE3 is expressed on a sulfoglycolipid in addition to neoplastic mucins. , 2011, Biochemical and biophysical research communications.

[2]  L. Sibley.  Invasion and intracellular survival by protozoan parasites. , 2011, Immunological reviews.

[3]  U. Krengel,et al.  Crystal structures exploring the origins of the broader specificity of escherichia coli heat-labile enterotoxin compared to cholera toxin. , 2010, Journal of molecular biology.

[4]  R. Cole,et al.  Toxoplasma gondii protease TgSUB1 is required for cell surface processing of micronemal adhesive complexes and efficient adhesion of tachyzoites , 2010, Cellular microbiology.

[5]  H. Yoshida,et al.  X-ray Structures of Human Galectin-9 C-terminal Domain in Complexes with a Biantennary Oligosaccharide and Sialyllactose* , 2010, The Journal of Biological Chemistry.

[6]  R. Schwarz,et al.  Binding of Toxoplasma gondii Glycosylphosphatidylinositols to Galectin-3 Is Required for Their Recognition by Macrophages* , 2010, The Journal of Biological Chemistry.

[7]  R. Levy,et al.  Intermolecular interactions in a 44 kDa interferon-receptor complex detected by asymmetric reverse-protonation and two-dimensional NOESY. , 2010, Biochemistry.

[8]  M. Williamson,et al.  Dimerisation of the UBA domain of p62 inhibits ubiquitin binding and regulates NF-kappaB signalling. , 2010, Journal of molecular biology.

[9]  M. Blackman,et al.  Members of a Novel Protein Family Containing Microneme Adhesive Repeat Domains Act as Sialic Acid-binding Lectins during Host Cell Invasion by Apicomplexan Parasites* , 2009, The Journal of Biological Chemistry.

[10]  S. Curry,et al.  Detailed insights from microarray and crystallographic studies into carbohydrate recognition by microneme protein 1 (MIC1) of Toxoplasma gondii , 2009, Protein Science.

[11]  A. Bax,et al.  TALOS+: a hybrid method for predicting protein backbone torsion angles from NMR chemical shifts , 2009, Journal of biomolecular NMR.

[12]  G. Kozlov,et al.  Atypical binding of the Swa2p UBA domain to ubiquitin. , 2009, Journal of molecular biology.

[13]  D. Roos,et al.  In Silico Identification of Specialized Secretory-Organelle Proteins in Apicomplexan Parasites and In Vivo Validation in Toxoplasma gondii , 2008, PloS one.

[14]  M. Blackman,et al.  Structural insights into microneme protein assembly reveal a new mode of EGF domain recognition , 2008, EMBO reports.

[15]  W. Gronwald,et al.  Structure of the leech protein saratin and characterization of its binding to collagen. , 2008, Journal of molecular biology.

[16]  D. Soldati-Favre,et al.  Molecular dissection of host cell invasion by the apicomplexans: the glideosome. , 2008, Parasite.

[17]  P. Simpson,et al.  Complete resonance assignment of the first and second apple domains of MIC4 from Toxoplasma gondii, using a new NMRView-based assignment aid , 2008, Biomolecular NMR assignments.

[18]  Jean-Michel Claverie,et al.  Phylogeny.fr: robust phylogenetic analysis for the non-specialist , 2008, Nucleic Acids Res..

[19]  Thilo Stehle,et al.  Structural basis of GM1 ganglioside recognition by simian virus 40 , 2008, Proceedings of the National Academy of Sciences.

[20]  Marie-France Carlier,et al.  Toxoplasma profilin is essential for host cell invasion and TLR11-dependent induction of an interleukin-12 response. , 2008, Cell host & microbe.

[21]  H. Ward,et al.  Cryptosporidium p30, a Galactose/N-Acetylgalactosamine-specific Lectin, Mediates Infection in Vitro* , 2007, Journal of Biological Chemistry.

[22]  L. Fielding.  NMR methods for the determination of protein–ligand dissociation constants , 2007 .

[23]  Ben M. Webb,et al.  Comparative Protein Structure Modeling Using MODELLER , 2007, Current protocols in protein science.

[24]  Joachim Mueller,et al.  VACCINATION WITH MICRONEME PROTEIN NCMIC4 INCREASES MORTALITY IN MICE INOCULATED WITH NEOSPORA CANINUM , 2007, The Journal of parasitology.

[25]  Anne Imberty,et al.  N-Glycolyl GM1 Ganglioside as a Receptor for Simian Virus 40 , 2007, Journal of Virology.

[26]  Aditi Gupta,et al.  Structure of the Malaria Antigen AMA1 in Complex with a Growth-Inhibitory Antibody , 2007, PLoS pathogens.

[27]  Philip G. Evans,et al.  Neoglycolipid probes prepared via oxime ligation for microarray analysis of oligosaccharide-protein interactions. , 2007, Chemistry & biology.

[28]  P. Brown,et al.  The Microneme Proteins EtMIC4 and EtMIC5 of Eimeria tenella Form a Novel, Ultra-high Molecular Mass Protein Complex That Binds Target Host Cells* , 2007, Journal of Biological Chemistry.

[29]  P. Simpson,et al.  Atomic resolution insight into host cell recognition by Toxoplasma gondii , 2007, The EMBO journal.

[30]  P. Simpson,et al.  High-level bacterial expression and purification of apicomplexan micronemal proteins for structural studies. , 2007, Protein and peptide letters.

[31]  J. Boothroyd,et al.  Pulling together: an integrated model of Toxoplasma cell invasion. , 2007, Current opinion in microbiology.

[32]  Michael Nilges,et al.  ARIA2: Automated NOE assignment and data integration in NMR structure calculation , 2007, Bioinform..

[33]  P. McEwan,et al.  Crystal structure of the factor XI zymogen reveals a pathway for transactivation , 2006, Nature Structural &Molecular Biology.

[34]  S. Gordon,et al.  Ligands for the β-Glucan Receptor, Dectin-1, Assigned Using “Designer” Microarrays of Oligosaccharide Probes (Neoglycolipids) Generated from Glucan Polysaccharides* , 2006, Journal of Biological Chemistry.

[35]  W. Petri,et al.  Roles for the galactose-/N-acetylgalactosamine-binding lectin of Entamoeba in parasite virulence and differentiation. , 2005, Glycobiology.

[36]  E. Cota,et al.  A Novel Galectin-like Domain from Toxoplasma gondii Micronemal Protein 1 Assists the Folding, Assembly, and Transport of a Cell Adhesion Complex* , 2005, Journal of Biological Chemistry.

[37]  B Dufour,et al.  Foodborne infections in France. , 2005, Foodborne pathogens and disease.

[38]  R. Klingebiel,et al.  Primary cerebral toxoplasmosis: a rare case of ventriculitis and hydrocephalus in AIDS. , 2005, Clinical neuropathology.

[39]  H. Vial,et al.  Synergistic role of micronemal proteins in Toxoplasma gondii virulence , 2005, The Journal of experimental medicine.

[40]  J. Wastling,et al.  Identification and Characterization of a Neospora caninum Microneme-Associated Protein (NcMIC4) That Exhibits Unique Lactose-Binding Properties , 2004, Infection and Immunity.

[41]  Ten Feizi,et al.  Oligosaccharide microarrays to decipher the glyco code , 2004, Nature Reviews Molecular Cell Biology.

[42]  M. Nilges,et al.  Refinement of protein structures in explicit solvent , 2003, Proteins.

[43]  C. Dominguez,et al.  HADDOCK: a protein-protein docking approach based on biochemical or biophysical information. , 2003, Journal of the American Chemical Society.

[44]  D. Hill,et al.  Toxoplasma gondii: transmission, diagnosis and prevention. , 2002, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[45]  M. Bienz,et al.  Identification of a Neospora caninum Microneme Protein (NcMIC1) Which Interacts with Sulfated Host Cell Surface Glycosaminoglycans , 2002, Infection and Immunity.

[46]  C. Roodveldt,et al.  Trypanosoma cruzi: identification of a galactose-binding protein that binds to cell surface of human erythrocytes and is involved in cell invasion by the parasite. , 2002, Experimental parasitology.

[47]  V. Carruthers.  Host cell invasion by the opportunistic pathogen Toxoplasma gondii. , 2002, Acta tropica.

[48]  T L Blundell,et al.  Crystal structures of NK1–heparin complexes reveal the basis for NK1 activity and enable engineering of potent agonists of the MET receptor , 2001, The EMBO journal.

[49]  S. R. Pereira,et al.  Toxoplasma gondii micronemal protein MIC1 is a lactose-binding lectin. , 2001, Glycobiology.

[50]  A. Otto,et al.  Characterization and crystallization of a novel Sarcocystis muris lectin, SML-2. , 2001, Acta crystallographica. Section D, Biological crystallography.

[51]  L. Sibley,et al.  The Toxoplasma Micronemal Protein MIC4 Is an Adhesin Composed of Six Conserved Apple Domains* , 2001, The Journal of Biological Chemistry.

[52]  D. Soldati,et al.  Identification and Characterization of an Escorter for Two Secretory Adhesins in Toxoplasma gondii , 2001, The Journal of cell biology.

[53]  D. Ferguson,et al.  The microneme protein MIC4, or an MIC4-like protein, is expressed within the macrogamete and associated with oocyst wall formation in Toxoplasma gondii. , 2000, International journal for parasitology.

[54]  D. Soldati,et al.  A dibasic motif in the tail of a class XIV apicomplexan myosin is an essential determinant of plasma membrane localization. , 2000, Molecular biology of the cell.

[55]  C. Hedberg,et al.  Food-related illness and death in the United States. , 1999, Emerging infectious diseases.

[56]  John E. Coligan,et al.  Current Protocols in Protein Science , 1996 .

[57]  J. Zimmerberg,et al.  Toxoplasma invasion: the parasitophorous vacuole is formed from host cell plasma membrane and pinches off via a fission pore. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[58]  J. Kovacs,et al.  Preventing toxoplasmic encephalitis in persons infected with human immunodeficiency virus. , 1995, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[59]  T. Wellems,et al.  Evidence for a switching mechanism in the invasion of erythrocytes by Plasmodium falciparum. , 1990, The Journal of clinical investigation.

[60]  A. Brini.  [Ocular manifestations of toxoplasmosis]. , 1958, Strasbourg medical.

[61]  Anne Imberty,et al.  N-Glycolyl GM 1 Ganglioside as a Receptor for Simian Virus 40 † , 2007 .

[62]  E. Meeusen,et al.  Galectins in parasite infection and allergic inflammation , 2004, Glycoconjugate Journal.

[63]  I. Campbell,et al.  Solution structure of a PAN module from the apicomplexan parasite Eimeria tenella , 2004, Journal of Structural and Functional Genomics.

[64]  Lee Fielding,et al.  NMR methods for the determination of protein-ligand dissociation constants. , 2003, Current topics in medicinal chemistry.