Abstract
Entamoeba histolytica is the causative agent of amoebiasis, and Entamoeba dispar is its noninvasive morphological twin. Entamoeba invadens is a reptilian parasite. In the present study, Western blot, phosphatase activity, immunofluorescence, and bioinformatic analyses were used to identify PP2C phosphatases of E. histolytica, E. dispar, and E. invadens. PP2C was identified in trophozoites of all Entamoeba species and cysts of E. invadens. Immunoblotting using a Leishmania mexicana anti-PP2C antibody recognized a 45.2 kDa PP2C in all species. In E. histolytica and E. invadens, a high molecular weight element PP2C at 75 kDa was recognized, mainly in cysts of E. invadens. Immunofluorescence demonstrated the presence of PP2C in membrane and vesicular structures in the cytosol of all species analyzed. The ~75 kDa PP2C of Entamoeba spp. shows the conserved domain characteristic of phosphatase enzymes (according to in silico analysis). Possible PP2C participation in the encystation process was discussed.
Highlights
IntroductionEntamoeba histolytica is an anaerobic parasitic amoebozoan and the causal agent of amoebiasis, which manifests as gastrointestinal disorders (e.g., amoebic colitis and dysentery) and less commonly as extraintestinal ulcers (e.g., amoebic liver abscesses, purulent pericarditis, and cerebral amoebiasis) [1,2,3]
Entamoeba histolytica is an anaerobic parasitic amoebozoan and the causal agent of amoebiasis, which manifests as gastrointestinal disorders and less commonly as extraintestinal ulcers [1,2,3]
E. histolytica trophozoites (HM1: IMSS) were cultured according to the Diamond Method [25], while E. dispar (SAW 760 strain) and E. invadens (IP-1; ATCC 30994) trophozoites were maintained in LYI-S-2 medium TYI-S-33 and incubated at 37°C and 28°C, respectively [26]
Summary
Entamoeba histolytica is an anaerobic parasitic amoebozoan and the causal agent of amoebiasis, which manifests as gastrointestinal disorders (e.g., amoebic colitis and dysentery) and less commonly as extraintestinal ulcers (e.g., amoebic liver abscesses, purulent pericarditis, and cerebral amoebiasis) [1,2,3]. E. histolytica has a faecal-oral life cycle involving a trophozoite stage and a cyst stage, the first of which is motile and proliferative, while the second is infective. The cysts enter humans by means of infected food or water and pass through the digestive tract until reaching the ileum. There, cysts undergo the process of excystation, consisting of the emergence of immature trophozoites from the cysts and their migration towards the colon, where they mature and proliferate [5, 6]. E. histolytica does not form cysts in vitro.
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