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Nicastrin (NICT) is a transmembrane protein physically associated with the polytypical aspartyl protease presenilin that plays a vital role in the correct localization and stabilization of presenilin to the membrane-bound γ-secretase complex. This complex is involved in the regulation of a wide range of cellular events, including cell signaling and the regulation of endocytosed membrane proteins for their trafficking and protein processing. Methods: In <i>Trypanosoma cruzi</i>, the causal agent of the Chagas disease, a NICT-like protein (Tc/NICT) was identified with a short C-terminus orthologous to the human protein, a large ectodomain (ECD) with numerous glycosylation sites and a single-core transmembrane domain containing a putative TM-domain (457GSVGA461) important for the γ-secretase complex activity. Results: Using the Spot-synthesis strategy with Chagasic patient sera, five extracellular epitopes were identified and synthetic forms were used to generate rabbit anti-Tc/NICT polyclonal serum that recognized a ~72-kDa molecule in immunoblots of <i>T. cruzi</i> epimastigote extracts. Confocal microscopy suggests that Tc/NICT is localized in the flagellar pocket, which is consistent with data from our previous studies with a <i>T. cruzi</i> presenilin-like protein. Phylogenetically, Tc/NICT was localized within a subgroup with the <i>T. rangeli</i> protein that is clearly detached from the other Trypanosomatidae, such as <i>T. brucei.</i> These results, together with a comparative analysis of the selected peptide sequence regions between the <i>T. cruzi</i> and mammalian proteins, suggest a divergence from the human NICT that might be relevant to Chagas disease pathology. As a whole, our data show that a NICT-like protein is expressed in the infective and replicative stages of <i>T. cruzi</i> and may be considered further evidence for a <b>γ</b>-secretase complex in trypanosomatids.