title: Purification and functional characterization of the 112 kDa adhesin of Entamoeba histolytica.
abstract: The 112 kDa adhesin of E. histolytica is directly involved in the cytopathogenic activity of the parasite. We describe here the purification of the 112 kDa protein by electroelution and immunoaffinity chromatography using a monoclonal antibody against the adhesin. Two proteins of 70 and 50 kDa were eluted from the immunoaffinity column along with the 112 kDa adhesin. The three proteins were recognized by monospecific polyclonal antibodies against the adhesin. The same peptides (72 and 56 kDa) were also observed after incubation of the purified intact adhesin in diethylamine buffer. Proteins of 112 and 72 kDa were found to have protease activity, evidenced by their ability to degrade gelatin. Our results indicate that the 112 kDa adhesin was specifically broken down into two polypeptides of 50-56 and 70-72 kDa. The significance of this in vivo is as yet unclear. The adhesin has proteolytic activity, which is retained in the 70-72 kDa polypeptide but not in the 50-56 kDa one.
authors.affiliation: Parasitology Laboratory, Faculty of Pharmacy, University of Paris, Chatenay-Malabry, France.
authors.forename: M C
authors.initials: MC
authors.lastname: Rigothier
authors.forename: G
authors.initials: G
authors.lastname: García-Rivera
authors.forename: M
authors.initials: M
authors.lastname: Guaderrama
authors.forename: E
authors.initials: E
authors.lastname: Orozco
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology; D000918:Antibody Specificity; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004748:Entamoeba histolytica / Q000737:chemistry* / Q000472:pathogenicity; D007163:Immunosorbent Techniques; D037102:Lectins; D008562:Membrane Glycoproteins; D010446:Peptide Fragments / Q000378:metabolism; D015800:Protozoan Proteins; D013379:Substrate Specificity; D014774:Virulence
chemical_list: D000913:Antibodies, Protozoan; D037102:Lectins; D008562:Membrane Glycoproteins; D010446:Peptide Fragments; D015800:Protozoan Proteins; C052452:galactose inhibitable adherence protein, Entamoeba histolytica