Dataset Information

Quantitative proteomic analysis of a Toxoplasma gondii mutant of the ABCB7 mitochondrial transporter

ABSTRACT: Toxoplasma gondii is a parasitic protist that is the agent of toxoplasmosis. It is capable of infecting a wide variety of vertebrates, including humans. The infection is mainly asymptomatic in immunocompetent patients, but in case of immunosuppression or for the congenital form of toxoplasmosis it can lead to severe pathologies with a possible fatal outcome. Like for other eukaryotes, many key cellular functions in T. gondii involve proteins containing an iron-sulfur cluster as a cofactor. Cytosolic and nuclear iron-sulfur proteins depend on a specific pathway for assembling their iron-sulfur cofactor. It was demonstrated in other eukaryotes (ie in the budding yeast model) that a sulfur-containing precursor originating from the mitochondrion and transported through the ABCB7 transporter is essential for building cytosolic iron-sulfur clusters. We have investigated the T. gondii homolog of the ABCB7 transporter by generating a specific mutant on which we performed a quantitative proteomic analysis to get insights into its involvement in the biogenesis of cytosolic and nuclear iron-sulfur proteins.

INSTRUMENT(S): Orbitrap Exploris 240

ORGANISM(S): Toxoplasma Gondii

DISEASE(S): Toxoplasmosis

SUBMITTER: valerie Rofidal

LAB HEAD: Sébastien Besteiro

PROVIDER: PXD048386 | Pride | 2024-08-09

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	PF_220413_ToxoFeS_A-Parent-1.raw	Raw
	PF_220413_ToxoFeS_A-Parent-2.raw	Raw
	PF_220413_ToxoFeS_A-Parent-3.raw	Raw
	PF_220413_ToxoFeS_A-rABCB7-1.raw	Raw
	PF_220413_ToxoFeS_A-rABCB7-2.raw	Raw

Items per page:

1 - 5 of 27

Similar Datasets

Project description:Tymoshenko2015 - Genome scale metabolic model - ToxoNet1 This model is described in the article: Metabolic Needs and Capabilities of Toxoplasma gondii through Combined Computational and Experimental Analysis. Tymoshenko S, Oppenheim RD, Agren R, Nielsen J, Soldati-Favre D, Hatzimanikatis V. PLoS Comput. Biol. 2015 May; 11(5): e1004261 Abstract: Toxoplasma gondii is a human pathogen prevalent worldwide that poses a challenging and unmet need for novel treatment of toxoplasmosis. Using a semi-automated reconstruction algorithm, we reconstructed a genome-scale metabolic model, ToxoNet1. The reconstruction process and flux-balance analysis of the model offer a systematic overview of the metabolic capabilities of this parasite. Using ToxoNet1 we have identified significant gaps in the current knowledge of Toxoplasma metabolic pathways and have clarified its minimal nutritional requirements for replication. By probing the model via metabolic tasks, we have further defined sets of alternative precursors necessary for parasite growth. Within a human host cell environment, ToxoNet1 predicts a minimal set of 53 enzyme-coding genes and 76 reactions to be essential for parasite replication. Double-gene-essentiality analysis identified 20 pairs of genes for which simultaneous deletion is deleterious. To validate several predictions of ToxoNet1 we have performed experimental analyses of cytosolic acetyl-CoA biosynthesis. ATP-citrate lyase and acetyl-CoA synthase were localised and their corresponding genes disrupted, establishing that each of these enzymes is dispensable for the growth of T. gondii, however together they make a synthetic lethal pair. This model is hosted on BioModels Database and identified by: MODEL1504280000. To cite BioModels Database, please use: BioModels Database: An enhanced, curated and annotated resource for published quantitative kinetic models. To the extent possible under law, all copyright and related or neighbouring rights to this encoded model have been dedicated to the public domain worldwide. Please refer to CC0 Public Domain Dedication for more information.

Project description:The mitochondrial ATP-binding cassette (ABC) transporters ABCB7 in humans, Atm1 in yeast and ATM3 in plants are highly conserved in their overall architecture and particularly in their glutathione binding pocket located within the transmembrane spanning domains. These transporters have attracted interest in the last two decades based on their proposed role in connecting the mitochondrial iron-sulfur (Fe-S) cluster assembly with its cytosolic Fe-S cluster assembly (CIA) counterpart. So far, the specific compound that is transported across the membrane remains unknown. In this report we characterized the ABCB7-like transporter Rcc02305 in Rhodobacter capsulatus, which shares 47% amino acid sequence identity to its mitochondrial counterpart. The constructed interposon mutant strain in R. capsulatus displayed increased levels of intracellular reactive oxygen species without a simultaneous accumulation of the cellular iron concentration. The inhibition of endogenous glutathione biosynthesis resulted in an increase of total glutathione levels in the mutant strain. Bioinformatic analysis of the amino acid sequence motifs revealed an aminotransferase class-V pyridoxal-5’-phosphate (PLP) binding site overlapping with the Walker A motif within the nucleotide binding domains of the transporter. The PLP cofactor is well characterized in L-cysteine desulfurases like IscS and NFS1 for its role in the formation of a protein-bound persulfide group. In this report, we present a model proposing that this ABC-transporter in R. capsulatus in its inward facing open conformation first produces a nucleophilic persulfide, which further reacts in a substitution reaction with a trapped GS(S)nG bound to the TMD resulting to a mixed glutathione polysulfide. In the next step, the release of the persulfide from the PLP binding site enables ATP to bind to the Walker A motif and the subsequent hydrolysis of ATP triggers a translocation of the mixed disulfide to the periplasm. We rename the ABCB7-like transporter Rcc02305 in R. capsulatus to PexA for PLP binding exporter.

Dataset Information

Quantitative proteomic analysis of a Toxoplasma gondii mutant of the ABCB7 mitochondrial transporter

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets