title: Proteomic identification of S-nitrosylated proteins in the parasite Entamoeba histolytica by resin-assisted capture: insights into the regulation of the Gal/GalNAc lectin by nitric oxide.
abstract: Entamoeba histolytica is a gastrointestinal protozoan parasite that causes amebiasis, a disease which has a worldwide distribution with substantial morbidity and mortality. Nitrosative stress, which is generated by innate immune cells, is one of the various environmental challenges that E. histolytica encounters during its life cycle. Although the effects of nitric oxide (NO) on the regulation of gene expression in this parasite have been previously investigated, our knowledge on S-nitrosylated proteins in E.histolytica is lacking. In order to fill this knowledge gap, we performed a large-scale detection of S-nitrosylated (SNO) proteins in E.histolytica trophozoites that were treated with the NO donor, S-nitrosocysteine by resin-assisted capture (RAC). We found that proteins involved in glycolysis, gluconeogenesis, translation, protein transport, and adherence to target cells such as the heavy subunit of Gal/GalNac lectin are among the S-nitrosylated proteins that were enriched by SNO-RAC. We also found that the S-nitrosylated cysteine residues in the carbohydrate recognition domain (CRD) of Gal/GalNAc lectin impairs its function and contributes to the inhibition of E.histolytica adherence to host cells. Collectively, these results advance our understanding of the mechanism of reduced E.histolytica adherence to mammalian cells by NO and emphasize the importance of NO as a regulator of key physiological functions in E.histolytica.
authors.affiliation: Department of Molecular Microbiology, The Bruce Rappaport Faculty of Medicine, Technion, Haifa, Israel.
authors.forename: Rivka
authors.initials: R
authors.lastname: Hertz
authors.affiliation: Department of Biochemistry, The Bruce Rappaport Faculty of Medicine, Technion, Haifa, Israel.
authors.forename: Shani
authors.initials: S
authors.lastname: Ben Lulu
authors.affiliation: Department of Molecular Microbiology, The Bruce Rappaport Faculty of Medicine, Technion, Haifa, Israel.
authors.forename: Preeti
authors.initials: P
authors.lastname: Shahi
authors.affiliation: Department of Molecular Microbiology, The Bruce Rappaport Faculty of Medicine, Technion, Haifa, Israel.
authors.forename: Meirav
authors.initials: M
authors.lastname: Trebicz-Geffen
authors.affiliation: Department of Biochemistry, The Bruce Rappaport Faculty of Medicine, Technion, Haifa, Israel.
authors.forename: Moran
authors.initials: M
authors.lastname: Benhar
authors.affiliation: Department of Molecular Microbiology, The Bruce Rappaport Faculty of Medicine, Technion, Haifa, Israel.
authors.forename: Serge
authors.initials: S
authors.lastname: Ankri
mesh_terms: D002241:Carbohydrates / Q000737:chemistry; D002448:Cell Adhesion; D002846:Chromatography, Affinity; D003545:Cysteine / Q000031:analogs & derivatives* / Q000737:chemistry; D004748:Entamoeba histolytica / Q000737:chemistry*; D004749:Entamoebiasis / Q000276:immunology / Q000469:parasitology; D006019:Glycolysis; D006367:HeLa Cells; D006801:Humans; D037102:Lectins / Q000737:chemistry*; D009569:Nitric Oxide / Q000737:chemistry*; D009584:Nitrogen / Q000737:chemistry*; D021381:Protein Transport; D020543:Proteome; D040901:Proteomics; D015800:Protozoan Proteins / Q000235:genetics; D026403:S-Nitrosothiols / Q000737:chemistry*
chemical_list: D002241:Carbohydrates; D037102:Lectins; D020543:Proteome; D015800:Protozoan Proteins; D026403:S-Nitrosothiols; D009569:Nitric Oxide; C021315:S-nitrosocysteine; D003545:Cysteine; D009584:Nitrogen