Identification and enzymatic activities of four protein disulfide isomerase (PDI) isoforms of Leishmania amazonensis

Hong, Baoyu; Soong, Lynn

doi:10.1007/s00436-007-0784-4

Cited by 31 publications

(34 citation statements)

References 41 publications

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“…Surprisingly, despite a high degree of homology with LmPDI (86% of identities and 96% of similarities), the 52-kDa L. amazonensis PDI protein displayed only the isomerase activity (Hong and Soong 2008). Moreover, in another Trypanosomatidae (T. brucei), the LmPDI orthologue, Td-PDI-2 also displays only the isomerase activity.…”

Section: Discussionmentioning

confidence: 99%

Leishmania major protein disulfide isomerase as a drug target

et al. 2011

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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.

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Section: Discussionmentioning

confidence: 99%

Leishmania major protein disulfide isomerase as a drug target

et al. 2011

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“…Four L. amazonensis PDIs encoding 52-, 47-, 40-, and 15 kDa proteins have been characterized. Homology analysis showed that the sequence identity between L. amazonensis (New World) PDIs and their counterpart PDI sequences from L. major (Old World) ranged from 76% to 99% [16]. Meek et al 2002 have detected IgG specific to four LaPDIs (especially to the 52 kDa PDI, known as PDI-2) [17].…”

Section: Protein Overexpression and Purificationmentioning

confidence: 99%

“…Taken together, these results demonstrate that the recombinant LtPDI-His is able to catalyze both oxidation and isomerization of protein disulfides. In a previous another study, Hong and Soong 2008 showed that the L. amazonensis 15, 40, and 47 kDa PDIs display both isomerase and reductase activities. Surprisingly, despite a high degree of homology with Lm PDI (86% of identities and 96% of similarities), the 52 kDa L. amazonensis PDI protein displayed only the isomerase activity [16].…”

Section: Pdi-2-stimulated Oxidation and Disulfide Bond Isomerization mentioning

confidence: 99%

“…In a previous another study, Hong and Soong 2008 showed that the L. amazonensis 15, 40, and 47 kDa PDIs display both isomerase and reductase activities. Surprisingly, despite a high degree of homology with Lm PDI (86% of identities and 96% of similarities), the 52 kDa L. amazonensis PDI protein displayed only the isomerase activity [16]. In the another study that was done by Padilla et al, 2003, the PDI-2 from L. donovani also displays both the oxidase and isomerase activities [23].…”

Section: Pdi-2-stimulated Oxidation and Disulfide Bond Isomerization mentioning

confidence: 99%

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Cloning, expression, purification and characterization of Leishmania tropica PDI-2 protein

et al. 2016

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In Leishmania species, protein disulfide isomerase (PDI) is an essential enzyme that catalyzes thiol-disulfide interchange. The present work describes the isolation, cloning, sequencing and expression of the pdI-2 gene. Initially, the gene was amplified from L. tropica genomic DNA by PCR using specific primers before cloning into the expression vector pET-15b. The construct pET/pdI-2 was transformed into BL21(DE3) cells and induced for the protein expression. SDS-PAGE and western blot analysis showed that the expressed protein is about 51 kDa. Cloned gene sequence analysis revealed that the deduced amino acid sequence showed significant homology with those of several parasites PDIs. Finally, recombinant protein was purified with a metal-chelating affinity column. The putative protein was confirmed as a thiol - disulfide oxidoreductase by detecting its activity in an oxidoreductase assay. Assay result of assay suggested that the PDI-2 protein is required for both oxidation and reduction of disulfide bonds in vitro. Antibodies reactive with this 51 kDa protein were detected by Western blot analysis in sera from human infected with L. tropica. This work describes for the first time the enzymatic activity of recombinant L. tropica PDI-2 protein and suggests a role for this protein as an antigen for the detection of leishmaniasis infection.

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“…Current evidence suggests that PDIs are also involved in a wide range of other biological functions in many cellular compartments including the cell surface, where they may participate in the reducing exterior environment and/or may be involved in cell adhesion (Turano et al, 2002). Parasite PDI-like enzymes were identified in Neospora caninum (Naguleswaran et al, 2005), Toxoplasma gondii (Meek et al, 2002a,b), Theileria parva (Ebel et al, 2002), different Leishmania species (Hong and Soong, 2008;Ben et al, 2002;Padilla et al, 2003), Cryptosporidium parvum (Blunt et al, 1996) and several species of Plasmodium (Florent et al, 2000;Mahajan et al, 2006;Mouray et al, 2007;Novo et al, 2009). T. gondii and N. caninum, both closely related to B. besnoiti (Ellis et al, 2000), have been shown to express highly homologous molecules, which could indicate that PDI plays a key role in the biology of neosporosis and toxoplasmosis (Ma et al, 2009;Meek et al, 2002a;Naguleswaran et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Besnoitia besnoiti protein disulfide isomerase (BbPDI): Molecular characterization, expression and in silico modelling

Marcelino

Martins

Morais

et al. 2011

Experimental Parasitology

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a b s t r a c tBesnoitia besnoiti is an apicomplexan parasite responsible for bovine besnoitiosis, a disease with a high prevalence in tropical and subtropical regions and re-emerging in Europe. Despite the great economical losses associated with besnoitiosis, this disease has been underestimated and poorly studied, and neither an effective therapy nor an efficacious vaccine is available. Protein disulfide isomerase (PDI) is an essential enzyme for the acquisition of the correct three-dimensional structure of proteins. Current evidence suggests that in Neospora caninum and Toxoplasma gondii, which are closely related to B. besnoiti, PDI play an important role in host cell invasion, is a relevant target for the host immune response, and represents a promising drug target and/or vaccine candidate. In this work, we present the nucleotide sequence of the B. besnoiti PDI gene. BbPDI belongs to the thioredoxin-like superfamily (cluster 00388) and is included in the PDI_a family (cluster defined cd02961) and the PDI_a_PDI_a 0 _c subfamily (cd02995). A 3D theoretical model was built by comparative homology using Swiss-Model server, using as a template the crystallographic deduced model of Tapasin-ERp57 (PDB code 3F8U chain C). Analysis of the phylogenetic tree for PDI within the phylum apicomplexa reinforces the close relationship among B. besnoiti, N. caninum and T. gondii. When subjected to a PDI-assay based on the polymerisation of reduced insulin, recombinant BbPDI expressed in E. coli exhibited enzymatic activity, which was inhibited by bacitracin. Antiserum directed against recombinant BbPDI reacted with PDI in Western blots and by immunofluorescence with B. besnoiti tachyzoites and bradyzoites.

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Identification and enzymatic activities of four protein disulfide isomerase (PDI) isoforms of Leishmania amazonensis

Cited by 31 publications

References 41 publications

Leishmania major protein disulfide isomerase as a drug target

Leishmania major protein disulfide isomerase as a drug target

Cloning, expression, purification and characterization of Leishmania tropica PDI-2 protein

Besnoitia besnoiti protein disulfide isomerase (BbPDI): Molecular characterization, expression and in silico modelling

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