Abstract
Leishmaniasis is a neglected tropical disease (NTD) caused by the protozoan parasite Leishmania. The disease has several clinical manifestations with visceral leishmaniasis (VL) being the most fatal. The apicomplexan protozoan has a digenetic life cycle between the sandfly vectors (promastigote stage) and the mammalian hosts (amastigote stage). Promastigote to amastigote differentiation is crucial for intracellular survival and for successful establishment of infection. However, the underlying molecular mechanisms still remain elusive. We investigated the proteome changes during the early differentiation stages of Leishmania donovani clinical isolate from an Amphotericin B (AmB) relapse patient. Sequential Window Acquisition of All Theoretical Mass Spectra (SWATH-MS) based quantitative proteomics revealed 814 and 921 differentially modulated proteins after 24 h and 48 h of promastigote to amastigote differentiation. We identified several key proteins like Tb-292-membrane-associated-protein-like-protein, Glycoprotein 96–92, p22 proteins, Maoc family dehydratase-like protein, KH domain-containing putative protein, Apolipoprotein-A1/A4/E-domain-containing-protein, Mkiaa0324 protein-like protein and Protein YIPF not reported hitherto. Several key metabolic proteins, proteases and proteins involved in post-translational modifications (PTM) were also differentially modulated between the amastigote and promastigote stages. Bioinformatics analyses of the proteomics data revealed several metabolic pathways viz., glycolysis, oxidative stress, fatty acid metabolism, asparagine/glutamine biosynthesis and purine/pyrimidine biosynthesis to be differentially regulated between the digenetic stages. Further studies to investigate the role of these proteins would allow better understanding of Leishmania pathogenesis and identification of targets for therapeutic intervention and biomarker studies. Data are available via ProteomeXchange with identifier PXD028775.
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30 May 2022
A Correction to this paper has been published: https://doi.org/10.1007/s42485-022-00089-y
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KP and SVK acknowledge the Seed grant from Savitribai Phule Pune University, Pune and PennState University, USA (SPPU–PennState University Joint MoU Research Fund). Funding to KP from UGC-CAS, DRDP, ICMR, DST-PURSE, and Government of India is also acknowledged.
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CBR: conceptualization, formal analysis, investigation, data curation, resource, writing—original draft. VC: data curation, DP: data curation, writing—original draft, RP: conceptualization, validation, writing—review & editing, SJ: data curation, SB: formal analysis, writing—review & editing, MJK: supervision, resources, writing—review & editing, SVK: resources, supervision, writing—review & editing, KP: conceptualization, resource, supervision, writing—review & editing.
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Supplementary file1 (DOCX 102 KB) Table 1 List of differentially expressed proteins identified in amastigotes after 24 h as compared to L. donovani promastigotes. Proteins with fold-change value ≥ 1.5 were considered as upregulated and proteins with fold-change value ≤ 0.6 were considered as down regulated proteins. Proteins with p value ≤0.05 were considered significant.
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Supplementary file2 (DOCX 116 KB) Table 2 List of differentially expressed proteins identified in amastigotes after 48 h as compared to L. donovani promastigotes. Proteins with fold-change value ≥ 1.5 were considered as upregulated and proteins with fold-change value ≤ 0.6 were considered as down regulated proteins. Proteins with p value ≤0.05 were considered significant.
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Routaray, C.B., Choudhari, V., Prakash, D. et al. Quantitative proteomic analysis reveals differential modulation of crucial stage specific proteins during promastigote to amastigote differentiation in Leishmania donovani. J Proteins Proteom 13, 17–27 (2022). https://doi.org/10.1007/s42485-021-00080-z
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DOI: https://doi.org/10.1007/s42485-021-00080-z