Chitinase secretion by encysting Entamoeba invadens and transfected Entamoeba histolytica trophozoites: Localization of secretory vesicles, endoplasmic reticulum, and Golgi apparatus

Entamoeba histolytica, the protozoan parasite that phagocytoses bacteria an d 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 vesi cles in encysting E. invadens parasites and in E. histolytica trophozoites overexpressing chitinase under an actin gene promoter. Abundant small secre tory vesicles were also identified with antibodies to the surface antigen A riel and with a fluorescent substrate of cysteine proteinases. Removal of a n 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 an d composed of a few large, perinuclear vesicles, was identified with antibo dies to ADP-ribosylating factor and to E-COP. We conclude that the amebic s ecretory pathway is similar to those of other eukaryotic cells, even if its appearance is somewhat different.