title: Characterization of cell surface carbohydrate receptors for Entamoeba histolytica adherence lectin.
abstract: Binding and cytolysis of Chinese hamster ovary (CHO) cells by Entamoeba histolytica trophozoites is inhibitable by galactose (Gal) or N-acetyl-D-galactosamine (GalNAc). To better define the carbohydrate receptor for E. histolytica, we compared the binding and cytolytic target properties of 10 CHO glycosylation mutants. Each mutant expresses a uniquely altered array of N- and/or O-linked cell surface carbohydrates. Amebic adherence was reduced when lactosamine-containing N-linked carbohydrates were essentially absent (Lec1 mutant), almost undetectable when Gal and GalNAc residues were absent on both N- and O-linked carbohydrates (ldlD.Lec1 mutant), and enhanced for mutants with increased terminal Gal residues (Lec2 and Lec3). Parental CHO cells treated with neuraminidase to expose Gal residues behaved like Lec2 mutants. Binding of purified Gal or GalNAc lectin to parental, Lec1, ldlD.Lec1, and Lec2 mutant CHO cells corroborated the adherence results. The suitability of CHO cell mutants as targets for amebic cytolysis correlated with their glycosylation phenotype: the Lec1 mutants were less susceptible than parental CHO cells, the ldlD.Lec1 mutants were highly resistant, and the Lec2 mutants required higher concentrations of Gal for inhibition. The E. histolytica Gal or GalNAc adherence lectin bound preferentially to beta 1-6-branched, N-linked carbohydrates lacking terminal sialic acid or fucose residues. However, amebic lectin binding to either N- or O-linked cell surface carbohydrates was sufficient to initiate parasite cytolytic activity.
authors.affiliation: Department of Medicine, University of Virginia School of Medicine, Charlottesville 22908.
authors.forename: J I
authors.initials: JI
authors.lastname: Ravdin
authors.forename: P
authors.initials: P
authors.lastname: Stanley
authors.forename: C F
authors.initials: CF
authors.lastname: Murphy
authors.forename: W A
authors.initials: WA
authors.lastname: Petri
mesh_terms: D000116:Acetylgalactosamine / Q000378:metabolism*; D000818:Animals; D002236:Carbohydrate Conformation; D002448:Cell Adhesion; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D003602:Cytotoxicity, Immunologic; D004748:Entamoeba histolytica / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D005260:Female; D005688:Galactosamine / Q000031:analogs & derivatives*; D005690:Galactose / Q000378:metabolism*; D006031:Glycosylation; D009154:Mutation; D010053:Ovary; D011971:Receptors, Immunologic / Q000032:analysis*; D011975:Receptors, Mitogen / Q000032:analysis*
chemical_list: D011971:Receptors, Immunologic; D011975:Receptors, Mitogen; C037045:galactose-N-acetylgalactosamine receptor; D005688:Galactosamine; D000116:Acetylgalactosamine; D005690:Galactose