title: Chitinase secretion by encysting Entamoeba invadens and transfected Entamoeba histolytica trophozoites: localization of secretory vesicles, endoplasmic reticulum, and Golgi apparatus.
abstract: Entamoeba histolytica, the protozoan parasite that phagocytoses bacteria and host cells, has a vesicle/vacuole-filled cytosol like that of macrophages. In contrast, the infectious cyst form has four nuclei and a chitin wall. Here, anti-chitinase antibodies identified hundreds of small secretory vesicles in encysting E. invadens parasites and in E. histolytica trophozoites overexpressing chitinase under an actin gene promoter. Abundant small secretory vesicles were also identified with antibodies to the surface antigen Ariel and with a fluorescent substrate of cysteine proteinases. Removal of an N-terminal signal sequence directed chitinase to the cytosol. Addition of a C-terminal KDEL peptide, identified on amebic BiP, retained chitinase in a putative endoplasmic reticulum, which was composed of a few vesicles of mixed sizes. A putative Golgi apparatus, which was Brefeldin A sensitive and composed of a few large, perinuclear vesicles, was identified with antibodies to ADP-ribosylating factor and to epsilon-COP. We conclude that the amebic secretory pathway is similar to those of other eukaryotic cells, even if its appearance is somewhat different.
authors.affiliation: Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, Massachusetts 02115, USA.
authors.forename: S K
authors.initials: SK
authors.lastname: Ghosh
authors.forename: J
authors.initials: J
authors.lastname: Field
authors.forename: M
authors.initials: M
authors.lastname: Frisardi
authors.forename: B
authors.initials: B
authors.lastname: Rosenthal
authors.forename: Z
authors.initials: Z
authors.lastname: Mai
authors.forename: R
authors.initials: R
authors.lastname: Rogers
authors.forename: J
authors.initials: J
authors.lastname: Samuelson
mesh_terms: D020727:ADP-Ribosylation Factors; D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000378:metabolism*; D001483:Base Sequence; D020126:Brefeldin A / Q000494:pharmacology; D002688:Chitinases / Q000378:metabolism*; D020756:Coatomer Protein; D003546:Cysteine Endopeptidases / Q000378:metabolism; D016054:DNA, Protozoan; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D004747:Entamoeba / Q000378:metabolism*; D004748:Entamoeba histolytica / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D019204:GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D019319:Okadaic Acid / Q000494:pharmacology; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology
chemical_list: D000953:Antigens, Protozoan; D020756:Coatomer Protein; D016054:DNA, Protozoan; D004791:Enzyme Inhibitors; D008565:Membrane Proteins; D011500:Protein Synthesis Inhibitors; D019319:Okadaic Acid; D020126:Brefeldin A; D002688:Chitinases; D003546:Cysteine Endopeptidases; D019204:GTP-Binding Proteins; D020727:ADP-Ribosylation Factors