PXD028939-1

PXD028939 is an original dataset announced via ProteomeXchange.

Title	Analysis of SLP-76 interactomes of human primary CD4+ T cells in presence or absence of Dasatinib treatmentAnalysis of SLP-76 interactomes of human primary CD4+ T cells in presence or absence of Dasatinib treatment
Description	Using a CRISPR/Cas9-based approach, we engineered human primary CD4+ T cells in which the bait protein SLP76 was tagged with an affinity Twin-Strep-tag (OST). Through affinity purification coupled with quantitative mass spectrometry, this system allows determining the composition and dynamics of the SLP76 signalosome following T cell activation, and it can be used to assess the mechanisms of action of drugs targeting human T cell activation. Dasatinib is an inhibitor that blocks the adenosine triphosphate binding sites of LCK. Here, we assessed the effect of the Dasatinib treatment on the SLP76 signalosome in CD4+ human T cells. Affinity purification of the OST tagged protein was performed using Streptactin beads, from T cells left non-stimulated, or stimulated for 30s or 120s with anti-CD3 and anti-CD28 antibodies. Prior to stimulation, they were preincubated for 45 min at 37°C with either Dasatinib (100 nM) or vehicle alone (DMSO). Each AP-MS purification was associated with a corresponding control (purification from non-edited WT CD4+ T cells, cultured and stimulated in the same conditions). The number of replicate biological experiments was n=4 for all conditions.
HostingRepository	PRIDE
AnnounceDate	2022-01-28
AnnouncementXML	Submission_2022-01-28_01:33:39.168.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Anne Gonzalez de Peredo
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	phosphorylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2021-10-04 10:03:23	ID requested
⏵ 1	2022-01-28 01:33:39	announced

Dataset with its publication pending

submitter keyword: TCR, SLP76, interactome, primary CD4+ human T cell, OST tag, CRISPR/CAS9, AP-MS, Dasatinib

Odile Schiltz
contact affiliation	Institute of Pharmacology and Structural Biology, CNRS, Toulouse, France
contact email	odile.schiltz@ipbs.fr
lab head
Anne Gonzalez de Peredo
contact affiliation	CNRS/IPBS
contact email	gonzalez@ipbs.fr
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD028939
     1. Label: PRIDE project
     2. Name: Analysis of SLP-76 interactomes of human primary CD4+ T cells in presence or absence of Dasatinib treatmentAnalysis of SLP-76 interactomes of human primary CD4+ T cells in presence or absence of Dasatinib treatment