Dataset Information

Aspartyl Protease 5 matures virulence factors found at the host-parasite interface in Toxoplasma gondii

ABSTRACT: N-terminome analysis of WT and ASP5 KO Toxoplasma parasites.

INSTRUMENT(S): Bruker Daltonics instrument model, Q Exactive

ORGANISM(S): Toxoplasma Gondii Me49 Toxoplasma Gondii

TISSUE(S): Cell Suspension Culture

SUBMITTER: Laura Dagley

LAB HEAD: Christopher J Tonkin

PROVIDER: PXD008574 | Pride | 2018-10-15

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	P3164_WTHKOL_12_1_1_1_01_4653.d.zip	Other
	P3164_WTLKOH_12_1_1_1_01_4666.d.zip	Other
	P3164_WTLKOH_4_1_1_1_01_4658.d.zip	Other
	P3196_WTLKOH_1_1_3_1_01_4863.d.zip	Other
	P3308_ASP5_1_1_1.raw	Raw

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Publications

Aspartyl Protease 5 Matures Dense Granule Proteins That Reside at the Host-Parasite Interface in Toxoplasma gondii.

Coffey Michael J MJ Dagley Laura F LF Seizova Simona S Kapp Eugene A EA Infusini Giuseppe G Roos David S DS Boddey Justin A JA Webb Andrew I AI Tonkin Christopher J CJ

mBio 20181030 5

Toxoplasma gondii infects approximately 30% of the world's population, causing disease primarily during pregnancy and in individuals with weakened immune systems. Toxoplasma secretes and exports effector proteins that modulate the host during infection, and several of these proteins are processed by the Golgi-associated aspartyl protease 5 (ASP5). Here, we identify ASP5 substrates by selectively enriching N-terminally derived peptides from wild-type and Δasp5 parasites. We r ...[more]

PMID: 30377279

Similar Datasets

Project description:Toxoplasma gondii infects approximately 30% of the world's population, causing disease primarily during pregnancy and in individuals with weakened immune systems. Toxoplasma secretes and exports effector proteins that modulate the host during infection, and several of these proteins are processed by the Golgi-associated aspartyl protease 5 (ASP5). Here, we identify ASP5 substrates by selectively enriching N-terminally derived peptides from wild-type and ?asp5 parasites. We reveal more than 2,000 unique Toxoplasma N-terminal peptides, mapping to both natural N termini and protease cleavage sites. Several of these peptides mapped directly downstream of the characterized ASP5 cleavage site, arginine-arginine-leucine (RRL). We validate candidates as true ASP5 substrates, revealing they are not processed in parasites lacking ASP5 or in wild-type parasites following mutation of the motif from RRL to ARL. All identified ASP5 substrates are dense granule proteins, and interestingly, none appear to be exported, thus differing from the analogous system in related Plasmodium spp. Instead we show that the majority of substrates reside within the parasitophorous vacuole (PV), and its membrane (the PVM), including two kinases and one phosphatase. We show that genetic deletion of WNG2 leads to attenuation in a mouse model, suggesting that this putative kinase is a new virulence factor in Toxoplasma Collectively, these data constitute the first in-depth analyses of ASP5 substrates and shed new light on the role of ASP5 as a maturase of dense granule proteins during the Toxoplasma lytic cycle.IMPORTANCE Toxoplasma gondii is one of the most successful human parasites. Central to its success is the arsenal of virulence proteins introduced into the infected host cell. Several of these virulence proteins require direct maturation by the aspartyl protease ASP5, and all require ASP5 for translocation into the host cell, yet the true number of ASP5 substrates and complete repertoire of effectors is currently unknown. Here we selectively enrich N-terminally derived peptides using Terminal Amine Isotopic Labeling of Substrates (TAILS) and use quantitative proteomics to reveal novel ASP5 substrates. We identify, using two different enrichment techniques, new ASP5 substrates and their specific cleavage sites. ASP5 substrates include two kinases and one phosphatase that reside at the host-parasite interface, which are important for infection.