Human Proteome Dataset Hek293, k652 and HeLa cell lines
Ontology highlight
ABSTRACT:
INSTRUMENT(S): Q Exactive
ORGANISM(S): Homo Sapiens (human)
TISSUE(S): Cell Culture
SUBMITTER: 
Henrik Zauber
LAB HEAD: Prof. Dr. Matthias Selbach
PROVIDER: PXD002383 | Pride | 2015-07-08
REPOSITORIES: Pride
Dataset's files
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Project description:Triple Silac approach to analyze cellline specific protein expression. Hek293 (Heavy, Lys8 and Arg10), K526 (Medium, DMEM with Lys 4 and Arg6), HeLa (Light).
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Project description:We used quantitative mass spectrometry-based proteomics to unravel global nerve growth factor (NGF)-induced changes in protein abundance using the rat PC12 cell line. Cells were stimulated with NGF for 0, 24 and 48 h in a triple SILAC setup (Light: Lys0,Arg0; Medium: Lys4,Arg6; and Heavy: Lys8,Arg10). All experiments were performed as biological replicates.
2015-05-06 | PXD001478 | Pride
Project description:We used quantitative mass spectrometry-based proteomics to unravel global nerve growth factor (NGF)-induced TrkA signaling dynamics at the interactome, phosphoproteome and proteome level. A tetracycline-inducible system for TrkA expression was generated in the human neuroblastoma cell line, SH-SY5Y. TrkA-induced cells were stimulated with NGF for different time points to follow phosphoproteome, interactome and proteome changes on a temporal scale. In a triple SILAC setup (Light: Lys0,Arg0; Medium: Lys4,Arg6; and Heavy: Lys8,Arg10), the samples were stimulated with NGF as indicated. Phosphoproteome: 0, 10, 45 min and 0, 120, 120 min+cycloheximide. Interactome: 0, 5, 10 min. Proteome: 0, 24, 48 h. All experiments were performed as biological replicates.
2015-05-06 | PXD001115 | Pride
Project description:Given the role of SUMOylation in pathogenic infection, we wanted to evaluate the changes to the host SUMO-2 subproteome during Shigella infection. HeLa cells overexpressing TAP-SUMO-2 or TAP-empty were used in conjunction with SILAC (Stable Isotope Labeling of Amino acids in Culture) (Golebiowski et al, 2009, 2010). Cells were grown in Dulbecco’s modified Eagle’s medium except that L-arginine and L-lysine were replaced with stable isotope (SILAC) forms depending on the treatment. Medium was supplemented with 10% dialyzed fetal calf serum. SILAC experiment compared TAP-containing cells (Lys0 and Arg0) with invasive Shigella flexneri (M90T) infected TAP-SUMO-2-containing cells (4,4,5,5-D4-lysine, Lys4, and 13C6- arginine, Arg6) and TAP-SUMO-2-containing cells infected with (mxiD) non-invasive strain of Shigella (13C6 15N2-lysine, Lys8, and 13C6 15N4 -arginine, Arg10).
2018-10-19 | PXD001100 | Pride
Project description:We used quantitative mass spectrometry-based proteomics to unravel global changes in the phosphoproteome upon treatment of ASP14 cells (a Ewing Sarcoma cell line) according to the setup below. The combination of drugs were experimentally shown to infer synergy in terms of cell killing. Three experiments were performed using a triple SILAC setup (Light: Lys0,Arg0; Medium: Lys4,Arg6; and Heavy: Lys8,Arg10): Experiment 1: Light SILAC: Drug combination (90 nM Midostaurin + 20 nM BMS-754807) Medium SILAC: Midostaurin (90 nM) Heavy SILAC: 0.1% DMSO Experiment 2: Light SILAC: Drug combination (90 nM Midostaurin + 20 nM BMS-754807) Medium SILAC: Midostaurin (90 nM) Heavy SILAC: BMS-754807 (20 nM) Experiment 3: Light SILAC: Drug combination (90 nM Midostaurin + 20 nM BMS-754807) Medium SILAC: BMS-754807 (20 nM) Heavy SILAC: 0.1% DMSO ASP14 cells were starved by replacing complete SILAC medium with SILAC minimal medium without serum over night. Cells were treated with drugs as stated above for 2h; then they were stimulated with serum for 20 minutes and harvested thereafter.
2017-04-19 | PXD002765 | Pride
Project description:We applied quantitative mass spectrometry (MS)-based proteomics to study the roles of Cbl and Cbl-b in long-term signaling responses related to neurite outgrowth and differentiation of SH-SY5Y neuroblastoma cells. Using stable isotope labeling by amino acids in cell culture (SILAC) and tandem mass tag (TMT)-labeling in combination with off-line high-pH reversed-phase fractionation and LC-MS/MS we analyzed how Cbl and Cbl-b depletion by siRNA affected the proteome, phosphoproteome and ubiquitylome of the neuroblastoma cells. SILAC proteome SILAC (Light Arg0/Lys0, medium Arg6/Lys4, heavy Arg10/Lys8) SH-SY5Y cells were treated with Cbl and Cbl-b or control (GFP) siRNA for 72 hours. For combined stimulation with ligand cocktail (FGF-2, IGF-1 PDGF-BB, TGFα) cells were treated with ligands for 48 h. Samples were analyzed in triplicates with set-up as described below: Set-up 1, 3 replicates (R1-3): Light: siGFP, Heavy: siCbl/siCbl-b Set-up 2, 3 replicates (E1-3): Light: siGFP + ligand cocktail, Medium: siCbl/siCbl-b, Heavy: siCbl/siCbl-b + ligand cocktail TMT phosphoproteome and proteome SH-SY5Y cells were treated with Cbl and Cbl-b siRNA, control (GFP) siRNA or Retinoic acid (RA) for 24 hours. Samples were prepared in triplicates and labelled with TMT10-plex reagents according to the set-up below: TMT10-126: siGFP E1 TMT10-127N: siCbl/siCbl-b E1 TMT10-127C: Retinoic acid E1 TMT10-128N: siGFP E2 TMT10-128C: siCbl/siCbl-b E2 TMT10-129N: Retinoic acid E2 TMT10-129C: siGFP E3 TMT10-130N: siCbl/siCbl-b E3 TMT10-130C: Retinoic acid E3 TMT10-131: Mix of the 9 samples SILAC Ubiquitin pulldown SILAC (Light Arg0/Lys0, heavy Arg10/Lys8) SH-SY5Y cells were treated with Cbl and Cbl-b or control (GFP) siRNA for 24 hours. Samples were analyzed in duplicates with set-up as described below: Light: siGFP, Heavy: siCbl/siCbl-b
2021-03-30 | PXD020389 | Pride
Project description:We used quantitative mass spectrometry-based proteomics to unravel global nerve growth factor (NGF)-induced TrkA signaling dynamics at the interactome, phosphoproteome and proteome level. A tetracycline-inducible system for TrkA expression was generated in the human neuroblastoma cell line, SH-SY5Y. TrkA-induced cells were stimulated with NGF for different time points to follow phosphoproteome, interactome and proteome changes on a temporal scale. In a triple SILAC setup (Light: Lys0,Arg0; Medium: Lys4,Arg6; and Heavy: Lys8,Arg10), the samples were stimulated with NGF as indicated. Phosphoproteome: 0, 10, 45 min and 0, 120, 120 min+cycloheximide. Interactome: 0, 5, 10 min. Proteome: 0, 24, 48 h. All experiments were performed as biological replicates.
2017-03-31 | MSV000080834 | MassIVE
Project description:A detailed description of the sample processing is published (Grün et al., 2014 Cell Rep). Briefly, a super Silac Mix using Lys8 was generated and spiked into all samples as an internal reference. Total Lysate was fractionated using SDS-PAGES. Up to 16 fractions were collected per sample. Proteins were in gel digest using LysC only.
2015-07-09 | PXD002482 | Pride
Project description:We developed a pulse-chase method in where fully SILAC (heavy, medium-heavy and Light) labelled cells were pulsed with Azidohomoalanine (AHA) and then chased for different length of times. These experiments were performed with 0, 1, 2, 4, 8, 16 and 13 h of chase. Data from 3 biological replicates are provided. Each replicate contains data from 3 experiments (0, 1, 2 and 0, 4, 8 and 0, 16, 32 hours chase) in where the 0 h time point is always heavy labelled followed by the medium and light labelled time points. In addition, AHA p-c experiments were performed using only 0, 4 and 8 h chase times in combination with different inhibitor combinations (MG132, Actinomycin D and Wortmanninin + Bafilomycin A1) or control (DMSO). Also, a control enrichment data set was created. Here heavy SILAC labelled cells were pulsed with AHA before being mixed with light cells that were not pulsed with AHA. This was followed by the click reaction and enrichment of AHA containing proteins. Label-swap experiments were also performed. Finally, a SILAC p-c data set is provided. Here light cells were pulsed with heavy (Arg10, Lys8) amino acids and then split in two. One half of the cells was chased in medium (Arg6, Lys4) amino acids and the other part was directly frozen. Label-swap experiments and experiments using different pulse and chase times were also performed. All provided datasets are from mouse fibroblast cells (NIH 3T3).
2016-10-06 | PXD004929 | Pride
Project description:Intracellular trafficking pathways in eukaryotic cells are essential to maintain organelle identity and structure, and to regulate cell communication with its environment. Shigella flexneri invades and subverts the human colonic epithelium by the injection of virulence factors through a type 3 secretion system (T3SS). In this work we report the multiple effects of two S. flexneri effectors, IpaJ and VirA, which target small GTPases of the Arf and Rab families, consequently inhibiting several intracellular trafficking pathways. IpaJ and VirA induce large-scale impairment of host protein secretion and block the recycling of surface receptors. Moreover, these two effectors decrease clathrin-dependent and -independent endocytosis. Therefore, S. flexneri infection induces a global blockage of host cell intracellular transport, affecting the exchange between cells and their external environment. The combined action of these effectors disorganizes the epithelial cell polarity, disturbs epithelial barrier integrity, promotes multiple invasion events and enhances the pathogen capacity to penetrate into the colonic tissue in vivo.
2019-06-04 | PXD012291 | Pride