Molecular analysis of the Gal/GalNAc adhesin of Entamoeba histolytica.
Attachment of Entamoeba histolytica to colonic epithelium and a variety of other target cells is mediated by a galactose/N-acetyl D-galactosamine (Gal/GalNAc) inhibitable adhesin. Seven monoclonal antibodies specific for nonoverlapping epitopes of the 170 kDa subunit have been shown to have distinct effects on adherence. Four of these monoclonal antibodies inhibit or have no effect on amebic adherence while two others enhance amebic adherence. The epitopes recognized by these seven monoclonal antibodies have been mapped to the extracellular cysteine rich region of the 170 kDa subunit. The conformational nature of the epitopes was examined by testing monoclonal antibody reactivity with isolated regions of the 170 kDa subunit expressed as fusion proteins in E. coli and also with denatured native adhesin. These analyses suggested that three of monoclonal antibodies recognized conformational epitopes while the remaining four recognized linear epitopes. The mapping of these monoclonal antibodies have identified functionally important regions of the Gal/GalNAc adhesin and have also shown that recombinant Gal/GalNAc adhesin, when expressed in E.coli, retained at least some of its native conformation.
Mann
B J
BJ
Department of Internal Medicine, University of Virginia Health Sciences Center, Charlottesville 22908, USA. bjm2r@virginia.edu
Lockhart
L A
LA
D000116:Acetylgalactosamine / Q000737:chemistry*; D000478:Alkylation; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000953:Antigens, Protozoan / Q000737:chemistry / Q000276:immunology; D000954:Antigens, Surface / Q000737:chemistry / Q000276:immunology; D015153:Blotting, Western; D002448:Cell Adhesion; D004591:Electrophoresis, Polyacrylamide Gel; D004748:Entamoeba histolytica / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D004749:Entamoebiasis / Q000469:parasitology; D018604:Epitope Mapping; D005690:Galactose / Q000737:chemistry*; D006790:Host-Parasite Interactions / Q000502:physiology; D006801:Humans; D037102:Lectins; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000276:immunology; D015800:Protozoan Proteins / Q000737:chemistry* / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000737:chemistry
D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D000954:Antigens, Surface; D037102:Lectins; D008562:Membrane Glycoproteins; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; C052452:galactose inhibitable adherence protein, Entamoeba histolytica; D000116:Acetylgalactosamine; D005690:Galactose