PXD018056

PXD018056 is an original dataset announced via ProteomeXchange.

Title	Target identification of the novel Toxoplasma gondii rhoptry kinase RON13
Description	The Toxoplasma gondii rhoptry neck protein RON13 was first identified as a substrate of aspartyl protease 3 by Terminal Amine Isotopic Labelling of Substrates (TAILS8, Dogga et al. 2017). It harbours a kinase domain the activity of which is important for host cell invasion. To identify the targets of RON13, we engineered a transgenic T. gondii line in the RH ΔKu80 background exploiting the tetracycline repressor system (Meissner et al. 2001). Here, the change of promoter resulted in the overall downregulation of RON13 expression leading to parasites that display significantly diminished infection capability. This defect can be rescued by complementation with an active version of RON13 kinase. We have performed phosphopeptide enrichment and shotgun proteomics using the parental line and three mutant strains: RON13 knock-down (RON13-KD), RON13 knock-down complemented with wildtype RON13 as well as RON13 knock-down complemented with the catalytically dead kinase in which an essential aspartate was replaced by alanine. Comparison of the results between the parental line and RON13-KD allowed us to identify phosphosites that were differentially phosphorylated. Validation of those phosphosites was performed using the comparison between the two complemented lines with the active or the inactive versions of RON13.
HostingRepository	PRIDE
AnnounceDate	2021-05-13
AnnouncementXML	Submission_2021-07-20_00:26:25.364.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Chandra Ramakrishnan
SpeciesList	scientific name: Toxoplasma gondii RH; NCBI TaxID: 383379;
ModificationList	phosphorylated residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2020-03-13 09:46:22	ID requested
1	2021-05-13 04:14:31	announced
⏵ 2	2021-07-20 00:26:26	announced	2021-07-20: Updated publication reference for PubMed record(s): 34145271.

Lentini G, Ben Chaabene R, Vadas O, Ramakrishnan C, Mukherjee B, Mehta V, Lunghi M, Grossmann J, Maco B, Visentin R, Hehl AB, Korkhov VM, Soldati-Favre D, Structural insights into an atypical secretory pathway kinase crucial for Toxoplasma gondii invasion. Nat Commun, 12(1):3788(2021) [pubmed]

submitter keyword: Toxoplasma gondii, rhoptry, kinase, LC-MS/MS

Prof. Adrian B. Hehl
contact affiliation	Institute of Parasitology, University of Zurich
contact email	adrian.hehl@access.uzh.ch
lab head
Chandra Ramakrishnan
contact affiliation	University of Zurich
contact email	chandra.ramakrishnan@uzh.ch
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2021/05/PXD018056

PRIDE project URI

• PRIDE
  1. PXD018056
     1. Label: PRIDE project
     2. Name: Target identification of the novel Toxoplasma gondii rhoptry kinase RON13