Trypanosoma cruzi is an obligate intracellular parasite that is the causative agent for Chagas Disease, a chronic bloodborne infection endemic to the Americas. Inositol-tetrakisphosphate 1-kinase (ITPK1) has been described as an enzyme that may mediate an alternative lipid-independent inositol polyphosphate synthesis pathway through phosphorylation of inositol monophosphate and other intermediates. Bioinformatics research has identified hypothetical protein TcCLB.503885.50 as a potential T. cruzi ITPK1 (TcITPK1) homolog and is orthologous to other higher-order eukaryotic ITPK1. The ability of TcITPK1 to act as the mediator for this alternative pathway has been established through yeast complementation assays and SAX-HPLC analysis of radioactively-labeled inositol phosphate species in complemented yeast. Immunofluorescence assays of T. cruzi epimastigotes with endogenously tagged TcITPK1 localizes to the cytosol. Additionally, preliminary data demonstrates TcITPK1 may be essential to T. cruzi survival as multiple CRISPR/Cas9 knockout experiments have resulted in no viable in vitro parasites in the epimastigote stage.
