An amebic protein disulfide isomerase (PDI) complements the yeast PDI1 mutation but is unable to support cell viability under ER or thermal stress

In eukaryotic cells, protein disulfide isomerases (PDI) are oxidoreductases that catalyze the proper disulfide bond formation during protein folding. The pathobiology of the protozoan parasite Entamoeba histolytica, the causative agent of human amebiasis, depends on secretion of several virulence factors, such as pore‐forming peptides and cysteine proteinases. Although the native conformation of these factors is stabilized by disulfide bonds, there is little information regarding the molecular machinery involved in the oxidative folding of amebic proteins. Whereas testing gene function in their physiological background would be the most suitable approach, we have taken advantage of the cellular benefits offered by the yeast Saccharomyces cerevisiae (as a model of eukaryotic cell) to examine the functional role of an amebic PDI (EhPDI). As the yeast PDI homolog is essential for cell viability, a functional complementation assay was carried out to test the ability of EhPDI to circumvent the lethal phenotype of a yeast PDI1 mutant. Also, its proficiency under stressful conditions was explored by examining the survival outcome following endoplasmic reticulum (ER) stress induced by a reductant agent (DTT) or thermal stress promoted by a nonpermissive temperature (37 °C). Our results indicate that EhPDI is functionally active when physiological conditions are stable. Nonetheless, when conditions are stressful (e.g., by the accumulation of misfolded proteins in the ER compartment), its functionality is exceeded, suggesting an inability to prevent unfolding, suppress aggregation, or assist refolding of proteins. Despite the latter, our findings constitute the initial step toward determining the participation of EhPDI in cellular mechanisms related to protein homeostasis.