title: Evidence for a novel Entamoeba histolytica lectin activity that recognises carbohydrates present on ovalbumin.
abstract: Entamoeba histolytica, an intestinal amoeba that causes dysentery and liver abscesses, acquires nutrients by engulfing bacteria in the colonic lumen and phagocytoses apoptotic cells during tissue invasion. In preliminary studies to identify ligands that stimulate amoebic phagocytosis, we used ovalbumin immobilized on latex particles as a potential negative control protein. Surprisingly, ovalbumin strongly stimulated E. histolytica particle uptake. Experiments using highly purified ovalbumin confirmed the specificity of this finding. The mechanism of particle uptake was actin-dependent, and the Entamoeba phagosome marker amoebapore A localised to ovalbumin-bead containing vacuoles. The most well described amoebic receptor is a Gal/GalNAc-specific lectin, but d-galactose had no effect on ovalbumin-stimulated phagocytosis. Ovalbumin has a single N-glycosylation site (Asn(292)) and is modified with oligomannose and hybrid-type oligosaccharides. We used both trifluoromethanesulfonic acid and N-glycanase to deglycosylate ovalbumin and tested the effect. Both methods substantially reduced the stimulatory effect of ovalbumin. Biotinylated ovalbumin bound the surface of fixed E. histolytica trophozoites saturably; furthermore, denatured ovalbumin and native ovalbumin both specifically inhibited ovalbumin-biotin binding, but deglycosylated ovalbumin had no effect. Collectively, these data suggest that E. histolytica has a previously unrecognised surface lectin activity that binds to carbohydrates on ovalbumin and stimulates phagocytosis.
authors.affiliation: Cell and Molecular Biology Program, University of Vermont College of Medicine, Burlington, VT 05405, USA.
authors.forename: Bradley T
authors.initials: BT
authors.lastname: Heron
authors.forename: Adam
authors.initials: A
authors.lastname: Sateriale
authors.forename: Jose E
authors.initials: JE
authors.lastname: Teixeira
authors.forename: Christopher D
authors.initials: CD
authors.lastname: Huston
mesh_terms: D050260:Carbohydrate Metabolism; D004748:Entamoeba histolytica / Q000378:metabolism*; D006801:Humans; D007473:Ion Channels / Q000032:analysis; D037102:Lectins / Q000378:metabolism*; D008863:Microspheres; D010047:Ovalbumin / Q000378:metabolism*; D010587:Phagocytosis; D010588:Phagosomes / Q000737:chemistry; D011485:Protein Binding; D015800:Protozoan Proteins / Q000032:analysis
chemical_list: D007473:Ion Channels; D037102:Lectins; D015800:Protozoan Proteins; C041670:amoebapore proteins, protozoan; D010047:Ovalbumin
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