PXD044182

PXD044182 is an original dataset announced via ProteomeXchange.

Title	Quantitative proteomic measurement of the Ca2+-dependent excretory-secretory antigen secretome of Toxoplasma gondii
Description	Apicomplexan parasites discharge specialized organelles called rhoptries upon host cell contact to mediate invasion. The events that drive rhoptry discharge are poorly understood, yet essential to sustain the apicomplexan parasitic life cycle. Rhoptry discharge appears to depend on proteins secreted from another set of organelles called micronemes, which in Toxoplasma gondii includes MIC8 and the microneme-associated CRMP complex. Here, we examine the function of the microneme protein CLAMP, uncovering its essential role in rhoptry discharge. CLAMP forms a distinct complex with two other microneme proteins, the invasion-associated SPATR, and a previously uncharacterized protein we name CLAMP-linked invasion protein (CLIP). Using a quantitative mass spectrometry based approach to determine the calcium-dependent excretory-secretory antigen secretome (MS-ESA), we identify secretion of both known and previously unverified microneme proteins. Our MS-ESA assay demonstrates that depletion of CLAMP or CLIP limits secretion of other CLAMP complex members, while not affecting secretion of other microneme proteins. CLAMP-deficiency does not impact parasite adhesion, however, knockdown of any member of the CLAMP complex affects rhoptry discharge. Phylogenetic analysis suggests orthologs of the essential complex components, CLAMP and CLIP, are ubiquitous across apicomplexans. Nevertheless, SPATR, which appears to act as an accessory factor in Toxoplasma, is essential for invasion during Plasmodium falciparum blood stages. Our results reveal a new protein complex that mediates rhoptry discharge following host-cell contact.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_09:13:42.199.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD044182
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Dylan Valleau
SpeciesList	scientific name: Toxoplasma gondii GT1; NCBI TaxID: 507601;
ModificationList	TMT6plex-126 reporter+balance reagent acylated residue; methylthiolated residue; ubiquitination signature dipeptidyl lysine; phosphorylated residue; L-methionine removal; acetylated residue; monohydroxylated residue; deamidated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2023-07-28 06:44:10	ID requested
1	2023-10-30 07:36:17	announced
⏵ 2	2023-11-14 09:13:51	announced	2023-11-14: Updated project metadata.

10.6019/PXD044182;

Valleau D, Sidik SM, Godoy LC, Acevedo-S, á, nchez Y, Pasaje CFA, Huynh MH, Carruthers VB, Niles JC, Lourido S, A conserved complex of microneme proteins mediates rhoptry discharge in Toxoplasma. EMBO J, 42(23):e113155(2023) [pubmed]

submitter keyword: Rhoptry secretion., Calcium,Toxoplasma gondii, MS-ESA

Sebastian Lourido
contact affiliation	Whitehead Institute for Biomedical Research and Department of Biology, Massachusetts Institute of Biology, Cambridge, MA, USA
contact email	lourido@wi.mit.edu
lab head
Dylan Valleau
contact affiliation	Whitehead Institute for Biomedical Research
contact email	valleau@wi.mit.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD044182
     1. Label: PRIDE project
     2. Name: Quantitative proteomic measurement of the Ca2+-dependent excretory-secretory antigen secretome of Toxoplasma gondii