Approaches for the identification of potential excreted/secreted proteins of<i>Leishmania major</i>parasites

Chenik, Mehdi; Lakhal, Sami; Khalef, N Ben; Zribi, Lilia; Louzir, Hechmi; Dellagi, Koussay

doi:10.1017/s0031182005009546

Cited by 34 publications

(41 citation statements)

References 109 publications

(95 reference statements)

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“…It is expressed and secreted at both promastigote and amastigote stages (Brotherton et al 2010;Chenik et al 2006;McNicoll et al 2006;Paape et al 2010). Preliminary results show that L. major parasites deleted for the lmpdi gene are non-virulent in the leishmaniasis experimental BALB/c mouse model (Ben Achour- Chenik et al 2011).…”

Section: Discussionmentioning

confidence: 99%

“…We initially identified this protein as a potential virulence factor of Leishmania parasites (Ben Achour et al 2002;Chenik et al 2006). First, we show that LmPDI displays similar structural domain organization and functional activities as PDIs from higher eukaryotes.…”

Section: Introductionmentioning

confidence: 99%

“…Previous studies suggest that protozoan PDI may play an essential role in virulence (Ben Achour et al 2002;Chenik et al 2006;Padilla et al 2003;Ramos et al 2008). In prokaryotes, the involvement of Disulfide Bond proteins (DsB), the PDI analogue, was widely described in bacterial pathogenicity (Heras et al 2009).…”

Section: Introductionmentioning

confidence: 99%

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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%

Section: Introductionmentioning

confidence: 99%

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Leishmania major protein disulfide isomerase as a drug target

et al. 2011

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“…Immuno-proteomic approaches have been developed to identify new Leishmania proteins with distinct biological functions, such as new diagnostic markers, vaccine candidates, and/or potential drug targets (80) (81) (82) . The use of antileishmanial antibodies obtained from infected mammalian hosts contributed to the refinement of these analyses, by assisting in the identifi cation of antigens recognized by the immune system during active disease (82) (83) .…”

Section: The Application Of Immuno-proteomic Approaches To the Identimentioning

confidence: 99%

Recent updates and perspectives on approaches for the development of vaccines against visceral leishmaniasis

Duarte

Lage

Martins

et al. 2016

Rev. Soc. Bras. Med. Trop.

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Visceral leishmaniasis (VL) is one of the most important tropical diseases worldwide. Although chemotherapy has been widely used to treat this disease, problems related to the development of parasite resistance and side effects associated with the compounds used have been noted. Hence, alternative approaches for VL control are desirable. Some methods, such as vector control and culling of infected dogs, are insuffi ciently effective, with the latter not ethically recommended. The development of vaccines to prevent VL is a feasible and desirable measure for disease control; for example, some vaccines designed to protect dogs against VL have recently been brought to market. These vaccines are based on the combination of parasite fractions or recombinant proteins with adjuvants that are able to induce cellular immune responses; however, their partial effi cacy and the absence of a vaccine to protect against human leishmaniasis underline the need for characterization of new vaccine candidates. This review presents recent advances in control measures for VL based on vaccine development, describing extensively studied antigens, as well as new antigenic proteins recently identifi ed using immuno-proteomic techniques.

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“…One positive transformant harboring the N-terminus is exposed and able to react with substrate cysteines [14]. The PDI protein in pathogens that are important for human infections [15]. Four L. amazonensis PDIs encoding 52-, 47-, 40-, and 15 kDa proteins have been characterized.…”

Section: Protein Overexpression and Purificationmentioning

confidence: 99%

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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Approaches for the identification of potential excreted/secreted proteins ofLeishmania majorparasites

Cited by 34 publications

References 109 publications

Leishmania major protein disulfide isomerase as a drug target

Leishmania major protein disulfide isomerase as a drug target

Recent updates and perspectives on approaches for the development of vaccines against visceral leishmaniasis

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

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