Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Characterization of RBM10 interactome in unchallenged and MK1775-treated cells

ABSTRACT: Mapping of RBM10-interactome using GFP-trap purification of EGFP-RBM10 expressed in HEK293T cells before and after WEE1 inhibition by MK1775. HEK293T expressing EGFP only or EGFP-RBM10 fusion (2 main isoforms) were left untreated or treated with 300nM MK1775 for 4 hours prior to GFP-trap enrichment and mass spectrometry analysis.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Lung, Epithelial Cell

DISEASE(S): Lung Adenocarcinoma

SUBMITTER: Feras Machour

LAB HEAD: Nabieh Ayoub

PROVIDER: PXD040109 | Pride | 2024-07-16

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	GFP_A_SCX_HFX.raw	Raw
	GFP_B_SCX_HFX.raw	Raw
	GFP_C_SCX_HFX.raw	Raw
	Peptides-results.xlsx	Xlsx
	ProteinGroups-results.xlsx	Xlsx

Items per page:

1 - 5 of 13

Similar Datasets

In planta expression of human polyQ-expanded huntingtin fragment reveals mechanisms to prevent disease-related protein aggregation.

Project description:We overexpressed eGFP, eGFP-SPP and pmRFP-Q74 construct in HEK93 cells. We made immunoprecipitation of GFP. We compare the interactome of eGFP and eGFP-SPP.

2023-08-11 | PXD044408 | Pride

Identification of nutrient dependend PIK3C2B regulators

Project description:To identify factors responsible for the differential localization and activity of PIK3C2Bin the presence or absence of serum mitogens we took a quantitative functional proteomics approach based on stable isotope labeling of amino acids in cell culture (SILAC). Genome-engineered HEK293T cells endogenously expressing eGFP-PI3KC15were cultured in serum-containing or serum-deprived media containing heavy or light amino acids, and subjected to affinity capture using a GFP trap matrix and LC-MS/MS

2019-10-02 | PXD014533 | Pride

Identification of RNA targets of EB1 using RIP-seq in Drosophila ovaries

Project description:Identification of RNAs that bind to EB1-GFP using RNA-immunoprecipitation followed by sequencing. Ovaries from flies expressing EB1-GFP (and GFP alone as a control) were used to immunoprecipitate crosslinked RNA-EB1GFP complexes using GFP Trap beads. The experiment was performed with three biological replicates for each.

2021-07-24 | E-MTAB-9078 | biostudies-arrayexpress

PALB2 interactome in asynchronous human cells

Project description:Identification of interacting partners of the Partner and Localizer of BRCA2 (PALB2), essential regulator of DNA repair by homologous recombination. Interacting partners of PALB2 were purified by GFP pull down from asynchronous HEK293 Flp-In T-REx cells, exogenously expressing Flag-EGFP tagged PALB2 at a similar level than the endogenous PALB2 level. Before GFP pull down, whole cell protein extracts were pre-cleared on IgG agarose beads to decrease binding of non-specific proteins during GFP pull down. GFP pull down were performed using GFP-Trap agarose beads (Chromotek) and washed at low salt concentration (150mM NaCl), to maintain interactions of low affinity binding protein partners. As a negative control, Flag-EGFP interacting proteins were purified, following the same protocol as described for the purification of Flag-EGFP PALB2 interacting proteins. Experiments were performed in triplicate.

2017-07-18 | PXD006391 | Pride

4SU-iCLIP of PTBP1 from HEK293 cells with or without MATR3 depletion

Project description:The aim of the experiment was to compare binding of PTBP1 in presence and absence of MATR3. HEK293T cells were transfected with siRNA targeting MATR3 or control sequences and labelled with 4thio-uridine for 8 hours (100µM, crosslinking 2x400mJ/cm2 365nm UV light).

2018-05-01 | E-MTAB-6286 | biostudies-arrayexpress

PALB2 ChAM acetylation mapping in human cells

Project description:The Chromatin Association Motif (ChAM) of the PArtner and Localizer of BRCA2 (PALB2) is essential for DNA repair function by homologous recombination in human cells. Molecular mechanisms governing ChAM interaction with chromatin remain unexplored. In this study, we have mapped acetylated lysines within ChAM domain, which are contained within a region essential to regulate its association with chromatin. GFP-ChAM sample submitted for proteomics analysis for acetylated lysines mapping was purified from HEK293T transiently expressing GFP-ChAM. GFP-ChAM was purified by GFP pull down using GFP-Trap agarose beads (Chromotek) and washed at 250mM NaCl.

2023-03-10 | PXD014678 | Pride

Analysis of interactions with truncated cytoplasmic dynein-2 intermediate chain WDR34-Q158*

Project description:Analysis of proteins interactiong with the trucated form of the cytoplasmic dynein-2 intermediate chain WDR34-Q158* using GFP-trap

2022-09-06 | PXD025686 | Pride

Identification of the interaction proteins of PRSS37 by GFP-IP/MS using testis proteins from PRSS37-EGFP knock-in mice and pAcr-EGFP transgenic mice

Project description:To identify the potential interaction partners of PRSS37, three testis lysates of Prss37E/+ mice (PRSS37-EGFP knock-in heterozygous mice) and three testis lysates of pAcr-EGFP transgenic mice were immunoprecipitated using anti-GFP mAb magnetic beads. The GFP-IP products of six samples from two groups were subsequently analyzed by mass spectrometry (MS) to identify differentially immmunoprecipitated proteins (DIPs) in the PRSS37-EGFP group. Proteins detected in pAcr-EGFP group were used as negative controls to exclude the endogenous proteins and polypeptides that interact nonspecifically with the anti-GFP mAb magnetic beads.

2021-05-06 | PXD024833 | Pride

MYO19 interacts weakly with Miro2 GTPases on the mitochondrial outer membrane

Project description:MYO19 interacts with mitochondria through a unique C-terminal mitochondrial association domain (MyMOMA). The specific molecular mechanisms for localization of MYO19 to mitochondria are poorly understood. Using promiscuous biotinylation, we have identified ~100 candidate MYO19 interacting proteins, a subset of which were also identified in via affinity-capture experiments. We chose to further explore the interaction between MYO19 and the mitochondrial GTPase Miro2 by expressing mchr-Miro2 in combination with GFP-tagged fragments of the MyMOMA domain and assaying for recruitment of MYO19-GFP to mitochondria. Co-expression of MYO19898-970-GFP with mchr-Miro2 enhanced MYO19898-970-GFP localization to mitochondria. Mislocalizing Miro2 to filopodial tips or the cytosolic face of the nuclear envelope did not recruit MYO19898-970-GFP to either location. To address the kinetics of the Miro2/MYO19 interaction, we used FRAP analysis and permeabilization-activated reduction in fluorescence (PARF) analysis. MyMOMA constructs containing a putative membrane insertion motif but lacking the complete Miro2 interacting region, MYO19853-935-GFP, displayed slow exchange kinetics. MYO19898-970-GFP, which does not include the membrane-insertion motif, displayed rapid exchange kinetics, suggesting that the MYO19 and Miro2 interaction is weaker than between MYO19 and the mitochondrial outer membrane. Mutation of well-conserved, charged residues within MYO19 or within the switch I and II regions of Miro2 abolished the enhancement of MYO19898-970-GFP localization in cells ectopically expressing mchr-Miro2. Additionally, expressing mutant versions of Miro2 thought to represent particular nucleotide states indicated that the enhancement of MYO19898-970-GFP localization is dependent on Miro2 nucleotide state. Taken together, these data suggest a role for Miro2 in regulation and integration of actin- and microtubule-based mitochondrial activities.

2019-09-10 | PXD015004 | Pride

Interactome of Irgm2 in mouse macrophages

Project description:This dataset corresponds to several experiments performed in order to identify proteins interacting with Immunity-related GTPase family M member 2 (Irgm2) in murine macrophages. To that aim, a version of the protein fused with the GFP (Egfp-Irgm2) was transduced in immortalized murine Bone Marrow-Derived Macrophages (iBMDMs), and Irgm2 complexes were subsequently purified using anti-GFP Trap technology. Different experiments were performed using either cells initially primed by treatment with interferon gamme (IFNg) or without any priming (NP), and in each case, cells were either infected with Salmonella (STm) or non infected (mock), leading to 4 different conditions. Corresponding control samples were prepared in the same way using macrophages transduced with GFP only (Egfp), leading to 8 different types of samples. Three independent biological experiments were performed for each condition, and each sample was analysed in triplicate by mass spectrometry, resulting in 72 LC-MS runs contained in the dataset.

2020-11-03 | PXD020457 | Pride