Leishmania major protein disulfide isomerase as a drug target

Abstract

Leishmaniasis is a major health problem worldwide and tools available for their control are limited. Effective vaccines are still lacking, drugs are toxic and expensive, and parasites develop resistance to chemotherapy. In this context, new antimicrobials are urgently needed to control the disease in both human and animal. Here, we report the enzymatic and functional characterization of a Leishmania virulence factor, Leishmania major Protein disulfide isomerase (LmPDI) that could constitute a potential drug target. LmPDI possesses domain structure organization similar to other PDI family members (a, a', b, b' and c domains), and it displays the three enzymatic and functional activities specific of PDI family members: isomerase, reductase and chaperone. These results suggest that LmPDI plays a key role in assisting Leishmania protein folding via its capacity to catalyze formation, breakage, and rearrangement of disulfide bonds in nascent polypeptides. Moreover, Bacitracin, a reductase activity inhibitor, and Ribostamycin, a chaperone activity inhibitor, were tested in LmPDI enzymatic assays and versus Leishmania promastigote in vitro cultures and Leishmania amastigote multiplication inside infected THP-1-derived macrophages. Bacitracin inhibited both isomerase and reductase activities, while Ribostamycin had no effect on the chaperone activity. Interestingly, Bacitracin blocked in vitro promastigote growth as well as amastigote multiplication inside macrophages with EC(50) values of 39 μM. These results suggest that LmPDI may constitute an interesting target for the development of new anti-Leishmania drugs.

Molecular characterization and immunolocalization of a protein disulfide isomerase from Angiostrongylus cantonensis

“…Our results showed that active enzyme can be obtained through heterologous expression and AcPDI was tested to behave oxidation-reduction activities as in other parasites (Mouray et al 2007;Hong and Soong 2008). These enzymatic activities were essential for the correct folding of nascent proteins and validate PDI as potential strategic targets for parasite diseases control (Epe et al 2007;Khalaf et al 2011). The co-expression of PDI and ancrod in Pichia pastoris can increase the production of ancrod, and the enzymatic activities of rAncrod were similar to that of the native protein (Zhang et al 2011).…”

Protein disulfide isomerase a multifunctional protein with multiple physiological roles

“…Historically the chaperone activity of PDI was assessed by a variety of methods depending on renaturation of denatured proteins monitored by activity-gain or loss-of-aggregates (Shao et al, 2000; Ben Khalaf et al, 2012; Wang et al, 2012; Hashimoto and Imaoka, 2013). A recent addition to this list utilizes acid-denatured green fluorescent protein (GFP), which interacts with a chaperone protein like PDI, and refold to yield the proper configuration and fluorescent properties (Mares et al, 2011).…”

Molecular Characterization and Analysis of a Novel Protein Disulfide Isomerase-Like Protein of Eimeria tenella

“…For instance, TgPDI of T. gondii is a novel vaccine candidate against toxoplasmosis because immunization with recombinant TgPDL elicits a significant protective immune reaction [57]. In Leishmania major , LmPDI is a virulence factor that is important in natural pathogenicity and might be a target for new anti-Leishmania drugs [20], [58]. Thus, EtPDIL might have unknown functions.…”