Dataset Information

Proteomic analysis of glycosomes from Leishmania donovani

ABSTRACT: Leishmania donovani is a kinetoplastid protozoan which causes Kala-azar or visceral leishmaniasis.Leishmania possess glycosomes that are unique and specialized subcellular microbody organelles. Glycosomes are known to harbor most peroxisomal enzymes and in addition they also possess glycolytic enzymes. In the present study, we carried out proteomic profiling of purified glycosomes isolated from L. donovani promastigotes using high resolution mass spectrometry. The majority of identified proteins are involved in metabolic processes such as carbohydrate, lipid and nucleic acid metabolism. Our present proteomic analysis is the most comprehensive study till date to map the proteome of L. donovaniglycosomes.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Leishmania Donovani

SUBMITTER: Akhilesh Pandey

LAB HEAD: AKHILESH PANDEY

PROVIDER: PXD000724 | Pride | 2015-03-25

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	LD_Glycosome_IG.pep.xml	Pepxml
	LD_Glycosome_IG.prot.xml	Xml
	LD_Glycosome_SCX.pep.xml	Pepxml
	LD_Glycosome_SCX.prot.xml	Xml
	Leish_Glyco_IG_1.raw	Raw

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Publications

Comprehensive proteomics analysis of glycosomes from Leishmania donovani.

Jamdhade Mahendra D MD Pawar Harsh H Chavan Sandip S Sathe Gajanan G Umasankar P K PK Mahale Kiran N KN Dixit Tanwi T Madugundu Anil K AK Prasad T S Keshava TS Gowda Harsha H Pandey Akhilesh A Patole Milind S MS

Omics : a journal of integrative biology 20150301 3

Leishmania donovani is a kinetoplastid protozoan that causes a severe and fatal disease kala-azar, or visceral leishmaniasis. L. donovani infects human host after the phlebotomine sandfly takes a blood meal and resides within the phagolysosome of infected macrophages. Previous studies on host-parasite interactions have not focused on Leishmania organelles and the role that they play in the survival of this parasite within macrophages. Leishmania possess glycosomes that are unique and specialized ...[more]

PMID: 25748437

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Project description:Infection with antimony resistant (SbR) but not with sentitive (SbS) Leishmania donovani (LD) gives rise to aggressive pathology in mammalian hosts, the cause of which is far from clear. Some intracellular pathogens exploit autophagy for their own benefit. Here we show that induction of autophagy in normal macrophages (MF) by pharmacological mediators prior to infection with SbRLD (SbRLD-MF) enhanced their growth as compared to untreated MF, unlike SbSLD-MF. Autophagy was evident in SbRLD-MF from electron microscopical studies showing double membrane-bound compartment around amastigote. In SbRLD-MF there is induction of beclin 1, which forms the platform to recruit other interacting molecules to initiate autophagy. Knocking down the beclin 1 transcription factor Nrf2 and subsequent infection with SbRLD showed significantly lower organ parasites as compared to wild type BALB/c mice. Cessation of autophagy in SbRLD-MF at the later stage of infection is coupled with induction of miR-30a, whose binding to 3'UTR of beclin 1 leads to its post-transcriptional attenuation followed by rise in intracellular Ca++ and apoptosis. SbRLD mediated translocation of AP-1 transcription factor to the nucleus induce pri-miR-30a over-expression. Rise in Ca++ causes caspase 8 activtion leading to the cleavage of beclin 1 and initiation of apoptosis in SbRLD-MF. Apoptosis may favor parasite egress for cell to cell transmission. We also found that beclin 1 expression is present in splenocytes of kala-azar patients harbouring SbRLD but not SbSLD. Our results suggest that SbRLD has evolved a unique mechanism for its own benefit which explains, in part, the cause of aggressive pathology. Peritoneal exudate macrophages were isolated from mouse, grown in 60mm plates and infected with Leishmania donovani and total RNA was isolated from cells at 12, 18 and 24 hrs post infection. Leishmania infected macrophage miRNA expression signature was generated. Cells grown on 60mm plates and infected with Leishmania. The main objective of the microarray analysis of mmu-miRNA in antimony resistant and antimony sensitive Leishmania donovani infected macrophages are as follows: 1. To study how the expression of miRNA varies in either antimony resistant or antimony sensitive Leishmania infected macrophages as compared to the normal macrophages as a function of time. LPS was used as control. 2. To study the expression of those miRNAs which are differentially expressed in antimony resistant and antimony sensitive Leishmania infected macrophages at each time point post infection. 3. To identify those miRNAs which are responsible for degradation of autophagy initiating protein beclin 1 mRNA