Human HEK293T nuclear extract LC-MSMS
Ontology highlight
ABSTRACT:
OTHER RELATED OMICS DATASETS IN: MODEL1109130000MODEL1909260003MODEL1909260004MODEL1703310000MODEL1909260005MODEL1909260006MODEL1311110000MODEL1311110001
INSTRUMENT(S): Q Exactive
ORGANISM(S): Homo Sapiens (human)
TISSUE(S): Epithelial Cell, Cell Culture
SUBMITTER: 
Chen Pu
LAB HEAD: Zhang Kai
PROVIDER: PXD014909 | Pride | 2020-01-13
REPOSITORIES: Pride
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| CP-BPTF-1-1_20190705.raw | Raw | |||
| CP-BPTF-1-2_20190705.raw | Raw | |||
| CP-BPTF-1-3_20190705.raw | Raw | |||
| CP-BPTF-2-1_20190705.raw | Raw | |||
| CP-BPTF-2-2_20190705.raw | Raw |
Items per page: 5 1 - 5 of 46 |
Similar Datasets
Project description:High-throughput phenotype-based screening of large libraries of novel compounds without known targets can identify small molecules that elicit a desired cellular response, but additional approaches are required to find and characterize their targets and mechanisms of action. Here we show that a compound termed lung cancer screen 3 (LCS3), previously selected for its ability to impair the growth of human lung adenocarcinoma (LUAD) cell lines, but not normal lung cells, induces oxidative stress and activates the NRF2 signaling pathway by generating reactive oxygen species (ROS) in sensitive LUAD cell lines. To identify the target that mediates this effect, we applied thermal proteome profiling (TPP) and uncovered the disulfide reductases GSR and TXNRD1 as LCS3 targets. Through enzymatic assays using purified protein, we confirmed that LCS3 inhibits disulfide reductase activity through a reversible, uncompetitive mechanism. Further, we demonstrate that LCS3-sensitive LUAD cells are correspondingly sensitive to the synergistic inhibition of glutathione and thioredoxin pathways. Lastly, a genome-wide CRISPR knockout screen identified the loss of NQO1 as a mechanism of LCS3 resistance. This work highlights the ability of TPP to uncover targets of small molecules identified by high-throughput screens and demonstrates the potential utility of inhibiting disulfide reductases as a therapeutic strategy for LUAD.
2022-02-23 | PXD027228 | Pride
Project description:Alternative 3’-end RNA processing provides an important source of transcriptome diversification, which impacts various biological processes and the etiology of diseases, including cancer. Here, we focused on a transcript isoform switch that leads to the readthrough expression of the long noncoding RNA NEAT1_2, at the expense of the shorter polyadenylated transcript NEAT1_1. Expression of NEAT1_2 is required for the formation of paraspeckles (PS), nuclear bodies that protect cancer cells from oncogene-induced replication stress and chemotherapy. Searching for proteins that interact with endogenous NEAT1 by RNA Affinity Purification coupled to mass spectrometry (RAP-MS), we identified factors involved in the 3’-end processing of polyadenylated (pA+) RNA as well as several components of the Integrator complex, known to process the 3’-ends of RNAs lacking polyadenylation sites. Unexpectedly, genetic perturbation experiments established that Integrator restraints NEAT1_2 expression, and thereby PS assembly, by promoting the 3’-end processing of pA+ NEAT1_1. Consistently, low expression levels of several Integrator subunits correlated with poorer prognosis of cancer patients exposed to chemotherapeutics. Our study identifies Integrator as a key regulator of PS biogenesis and highlights a previously unrecognized ability of the complex to process pA+ RNA. The data also establish a link between Integrator, cancer biology and chemosensitivity, which may be exploited therapeutically.
2020-10-14 | PXD015158 | Pride
Project description:Two-week old bread wheat seedlings hydroponically grown Hoagland solution were transferred to potassium (K+)-free conditions for 8 d, their root and leaf proteome profiles were assessed using iTRAQ proteome method, and NCBInr database combined with the recently published bread wheat genome information were used to analyze the identified protein species. Over 4,000 unique proteins were identified, 818 K+-responsive protein species showed significant abundance regulation. The most majority of the identified K+-responsive protein species had exact gene loci, and showed no global but tissue- and chromosome- dependent genome distributions. The identified protein species were associated with diverse functions and exhibited organ-specific differences. Most of identified protein species associated with hormone synthesis were the enzymes involved in the synthesis of jasmonic acid (JA). Allene oxide synthase (AOS), a key JA synthesis-related enzyme, was significantly induced in both root and leaf organs of K+-deficient wheat seedlings, and its overexpression in rice enhanced the tolerance to low K+ or K+ deficiency, increased contents of K+ and JA and transcription levels of some K+-responsive genes. However, rice AOS T-DNA inserted mutant (osaos) exhibited more sensitivity to K+ deficiency. K+ deficiency significantly increased abundance of a high affinity K+ transporter (TaAHAK1), TaAHAK1 transgenic rice seedlings markedly alleviated sensitivity to K+ deficiency, and K+ deficiency also upregulated expression of homologous OsHAK1 gene in TaAOS transgenic rice plants. These results suggested an essential role of JA in K+ deficiency and gave molecular insight into the responses of plant to K+ deficiency.
2017-10-09 | PXD003570 | Pride
Project description:During Agrobacterium rhizogenes–plant interaction, the rolB gene is transferred into the plant genome and is stably inherited in the plant’s offspring. Among the numerous effects of rolB on plant metabolism, including the activation of secondary metabolism, its effect on plant defense systems has not been sufficiently studied. Here, we performed a proteomic analysis of rolB-expressing Arabidopsis plants with a particular focus on defense proteins. In rolB plants, a reduced amount of scaffold proteins RACK1A, RACK1B, and RACK1C, known as receptors for activated C-kinase 1, was found. Proteomic analysis showed that rolB could potentially suppress the plant immune system by suppressing the RNA-binding proteins GRP7, CP29B, and CP31B, similar to the action of type III bacterial effectors. At the same time, rolB plants induce massive biosynthesis of protective proteins VSP1, VSP2, and PR4, which are markers of the activated jasmonate pathway. Moreover, rolB plants activate all components of the PYK10 defense complex of the endoplasmic reticulum involved in the metabolism of glucosinolates. We hypothesized that various defense systems activated by rolB are aimed at protecting the host plant from competing phytopathogens in order to create an effective ecological niche for A. rhizogenes. A RolB → RACK1A signaling module has been proposed that can exert most of the rolB-mediated effects on plant physiology and biochemistry.
2023-03-11 | PXD037959 | Pride
Project description:Canine heartworm is a widespread and potentially fatal mosquito-borne disease caused by infections with the parasitic nematode, Dirofilaria immitis. We have previously shown that systemic activation of the Toll immune pathway via silencing of the negative regulator Cactus in Aedes aegypti blocks parasite development in the Malpighian tubules, the mosquito renal organ. However, it was not established whether the Malpighian tubules were directly responding to Toll activation or were alternatively responding to upregulated proteins or other changes to the hemolymph driven by other tissues. Distinguishing these possibilities is crucial for developing more precise strategies to block D. immitis while potentially avoiding the fitness cost to the mosquito associated with Cactus silencing. This study defines the transcriptional response of Ae. aegypti Malpighian tubules after systemic Toll activation via intra-thoracic injection of dsCactus and found, like the response of whole mosquitoes, a significant increase in expression of Toll pathway target genes. Additionally, we identified a significant overlap between the transcriptional response of the Malpighian tubules and proteins upregulated in the hemolymph. Our data show that Malpighian tubules are capable of RNAi-mediated gene silencing and directly respond to dsCactus treatment by upregulating canonical Toll pathway targets. Though not definitive, the strong correspondence between the Malpighian tubule transcriptional and the hemolymph proteomic responses provides evidence that the tubules may contribute to mosquito humoral immunity.
2024-08-27 | PXD036225 | Pride
Project description:Diagnosis of ovarian cancer at an early stage is the most important determinant of survival. Thus, there is a clear need for novel biomarkers to improve diagnostic and prognostics that may better inform on therapeutic strategies. We have conducted a discovery study using label-free quantitative mass spectrometry (LFQ) to identify potential biomarker candidates in urine from individual ovarian cancer patients. LFQ analyses identified 4394 proteins (16397 peptides) in urine samples (n=20), 23 of which were significantly elevated in the malignant patient group compared to patients with benign disease. To validate these changes, we used Parallel Reaction Monitoring (PRM) to investigate their abundance in an independent cohort (n=20) of patient urine samples. Seven of the ten proteins were significantly enriched in the ovarian cancer patient samples; amongst these were established ovarian cancer markers WFDC2 (HE4) and Mesothelin (MSLN), validating our approach. This is the first application of a LFQ-PRM workflow to identify and validate ovarian cancer-specific biomarkers in urine samples.
2018-02-15 | PXD007962 | Pride
Project description:Nucleus as the important subcellular organ of plant cell is responsible for storing most genetic information and controlling the plant's activities. Seed germination is an important stage for plant growth. To understand the role of nuclear protein and its phosphorylation during rice seed germination, gel free/label free proteomic technique was used
2016-03-17 | PXD001774 | Pride
Project description:To investigate the repertoire of proteins interacting with the Aedes aegypti TUDOR protein AAEL012437 (Veneno), we transiently expressed a GFP-Veneno transgene in Aag2 cells. These cells were either infected with the alphavirus Sindbis virus (SINV) or mock-infected. Subsequently, cells were lysed and processed for immunoprecipitation using GFP-TRAP beads to purify Veneno-complexes. The same approach was followed for a truncated transgene lacking the first 205 amino acids (C206).
2018-12-12 | PXD009997 | Pride
Project description:As one of the most common maternal comorbidities, gestational diabetes mellitus (GDM) and preeclampsia (PE) are associated with maternal and infant health. Although the pathogenesis of PE and GDM remains controversial, oxidative stress is thought to be involved in the underlying pathology of GDM and PE. Protein lysine acetylation (Kac) plays an important regulatory role in biological processes. There is little data regarding the association of the maternal acetylome with GDM and PE. This study aimed to assess the multi-omics (proteome and acetylome) potential value in the underlying pathology for GDM and PE by label-free quantification proteomics technology. In our study, we included placental tissue from healthy individuals (n=6), GDM patients (n=6), and PE patients (n=6). We identified 22 overlapping DEPs (differentially expressed proteins) and 192 overlapping DAPs (differentially acetylated proteins) between the GDM and PE groups. Furthermore, 192 overlapping DAPs were mainly enriched in endoplasmic reticulum stress and ferroptosis pathways. Interestingly, endoplasmic reticulum stress, ferroptosis, and oxidative stress are believed to be related to each other. We also identified that acetylation of the HSP90AA1, HSPA8, PDIA3, GPX4, TF, and CP might serve as novel markers and better therapeutic targets in both complications. We thoroughly analyzed the key characteristics of proteome and acetylome in GDM and PE placental tissues, which may be useful for exploring the underlying pathology and discovering new biomarkers and therapeutic targets.
2024-01-25 | PXD032981 | Pride
Project description:Polarized vesicular trafficking directs specific receptors and ion channels to cilia, but the underlying mechanisms are poorly understood. Here we describe a role for DLG1, a core component of the Scribble polarity complex, in regulating ciliary protein trafficking in kidney epithelial cells. Conditional knockout of Dlg1 in mouse kidney caused causes ciliary elongation and cystogenesis, and cell-based proximity labelling proteomics and fluorescence microscopy showed alterations in the ciliary proteome upon loss of DLG1. Specifically, the retromer-associated protein SDCCAG3, IFT20 and polycystin-2 (PC2) were are reduced in cilia of DLG1 deficient cells compared to control cells. This phenotype was is recapitulated in vivo and rescuable by re-expression of wildtype DLG1, but not a Congenital Anomalies of the Kidney and Urinary Tract (CAKUT)-associated DLG1 variant, p.T489R. Finally, biochemical approaches and Alpha Fold modelling suggested that SDCCAG3 and IFT20 form a complex that associates, at least indirectly, with DLG1. Our work identifies a key role for DLG1 in regulating ciliary protein composition and suggests that ciliary dysfunction of the p.T489R DLG1 variant may contribute to CAKUT.
2024-05-14 | PXD051912 | Pride