Peptide fingerprinting of eluates obtained from phosphatase inhibitor beads (PIBs)
Ontology highlight
ABSTRACT:
INSTRUMENT(S): Orbitrap Exploris 480
ORGANISM(S): Homo Sapiens (human) Spodoptera Frugiperda
TISSUE(S): Cell Culture
SUBMITTER: 
WeiQiang Chen
LAB HEAD: Thomas Leonard
PROVIDER: PXD045891 | Pride | 2025-03-13
REPOSITORIES: Pride
Dataset's files
Items per page: 5 1 - 5 of 5 |
Similar Datasets
Project description:We aimed to identify proteins that would be turned over upon artificial tethering of WIPI2 to the mitochondrial surface upon rapalog treatment, using the FIS1-FRB plus FKBP-GFP-WIPI2 tethering system. We had previously observed that this induces mitophagy based on flow cytometry analysis of the mt-mKeima probe. Here, we aimed to identify which proteins are degraded upon 24 h Rapalog treatment and verified whether there was an enrichment of mitochondrial proteins being selectively turned over by mitophagy under these conditions in wild-type and NIX/BNIP3 double-knockout HeLa cells.
2025-01-30 | PXD060363 | Pride
Project description:We aimed to identify proteins that would be turned over upon DFP treatment, a small molecule that mimicks hypoxia treatment and which induces NIX/BNIP3 mitophagy. We aimed to identify which proteins are degraded upon 24 h DFP treatment and verified whether there was an enrichment of mitochondrial proteins being selectively turned over by mitophagy under these conditions.
2025-01-30 | PXD060356 | Pride
Project description:Proteomic expression analysis of Ca. Lokiarchaeum ossiferum, strain B35, grown using casein hydrolysate as the main carbon source under anaerobic conditions
2024-04-11 | PXD047696 | Pride
Project description:Staphylococcus aureus bacteria were decorated with modified ascorbate peroxidase APEX2-YFP-CWT via the cell wall targeting (CWT) domain of lysostaphin. Cytosol of transgenic HeLa cells expressing APEX2-YFP-CWT was generated by ultracentrifugation and living bacteria were incubated in the cytosol. The bacteria were stringently washed and added to either untreated human lung microvascular endothelial cells (HuLEC) or to cells pre-treated with bacterial sphingomyelinase (β-toxin), ionomycin, or amitriptyline. After 15 min, H2O2 was added for 1 min, then the biotinylation reaction was stopped by addition Stop Solution (containing sodium azide, sodium ascorbate, and Trolox in PBS). The cells then were scraped off the substratum, centrifuged, and lysed in RIPA buffer. To remove excess biotin phenol, lysates were transferred into a 3 kDa cutoff Amicon concentrator and 5 ml RIPA buffer was added. Solution was concentrated by centrifugation to 500-1000 µL. This step was repeated once. The lysates then were incubated overnight with magnetic streptavidin beads, followed by extensive washing and eventually biotinylated proteins were eluted by addition of Laemmli buffer containing an excess of biotin.
2025-03-13 | PXD051937 | Pride
Project description:Lysosomes serve dual antagonistic functions in cells by mediating anabolic growth signaling and the catabolic turnover of macromolecules. How these janus-faced activities are regulated in response to cellular nutrient status is poorly understood. We show here that lysosome morphology and function are reversibly controlled by a nutrient-regulated signaling lipid switch that triggers the conversion between motile mTOR complex 1 (mTORC1) signaling-active and static mTORC1-inactive degradative lysosomes clustered at the cell center. Starvation-triggered relocalization of phosphatidylinositol 4-phosphate [PI(4)P] metabolizing enzymes reshapes the lysosomal surface proteome to facilitate lysosomal proteolysis and to repress mTORC1-signaling by erasing phosphatidylinositol 3-phosphate [PI(3)P], which marks motile signaling-active lysosomes in the cell periphery. Interference with this PI(3)P/ PI(4)P lipid switch module impairs the adaptive response of cells to altering nutrient supply. Our data unravel a key function for lysosomal phosphoinositide metabolism in rewiring organellar membrane dynamics in response to cellular nutrient status.
2025-02-10 | PXD038428 | Pride
Project description:Non-alcoholic fatty liver disease (NAFLD) is gaining evermore importance due to rapidly rising incidence rates especially in western countries. As the disease progresses, it can develop into hepatocellular carcinoma. Related metabolic changes have been extensively studied in the past. However, the molecular mechanisms responsible for the development of fatty liver disease remain uncovered. Here we show that the basal phosphorylation of the MET receptor can be used as an indicator for hepatocyte dysregulation in fatty liver disease. Using primary hepatocytes isolated from a preclinical model fed with high-fat, high-sugar diet (Western diet; WD) or standard diet (SD), we find a strong downregulation of the PI3K-AKT pathway and upregulation of the MAPK pathway in the WD derived hepatocytes. By developing a mathematical model of HGF-induced signal transduction, calibrated with quantitative time-resolved measurements of both PI3K-AKT and MAPK pathways, we resolve molecular mechanisms responsible for these alterations. Thereby, we identify the basal MET phosphorylation rate as main driver for the altered signal transduction in WD hepatocytes that even leads to an increased proliferation behavior of the usually quiescent hepatocytes in the absence of growth factors. The adaptation of the dynamic pathway model of HGF signal transduction to patient-derived hepatocytes reveals a patient-specific variability of basal MET phosphorylation levels, which correlates with the clinical outcome of patients after liver surgery. These findings suggest that the dysregulated basal MET phosphorylation could be exploited to assess the health status of the liver and thereby inform estimation of the risk of a patient to suffer from liver failure after surgery.Non-alcoholic fatty liver disease (NAFLD) is gaining evermore importance due to rapidly rising incidence rates especially in western countries. As the disease progresses, it can develop into hepatocellular carcinoma. Related metabolic changes have been extensively studied in the past. However, the molecular mechanisms responsible for the development of fatty liver disease remain uncovered. Here we show that the basal phosphorylation of the MET receptor can be used as an indicator for hepatocyte dysregulation in fatty liver disease. Using primary hepatocytes isolated from a preclinical model fed with high-fat, high-sugar diet (Western diet; WD) or standard diet (SD), we find a strong downregulation of the PI3K-AKT pathway and upregulation of the MAPK pathway in the WD derived hepatocytes. By developing a mathematical model of HGF-induced signal transduction, calibrated with quantitative time-resolved measurements of both PI3K-AKT and MAPK pathways, we resolve molecular mechanisms responsible for these alterations. Thereby, we identify the basal MET phosphorylation rate as main driver for the altered signal transduction in WD hepatocytes that even leads to an increased proliferation behavior of the usually quiescent hepatocytes in the absence of growth factors. The adaptation of the dynamic pathway model of HGF signal transduction to patient-derived hepatocytes reveals a patient-specific variability of basal MET phosphorylation levels, which correlates with the clinical outcome of patients after liver surgery. These findings suggest that the dysregulated basal MET phosphorylation could be exploited to assess the health status of the liver and thereby inform estimation of the risk of a patient to suffer from liver failure after surgery.
2024-01-15 | PXD043007 | Pride
Project description:Data for the third YPIC Challenge. A synthetic peptide was analyzed using HR-MS. The peptide contains a codeword/phrase/sentence. Can you decrypt it? Get your best mates and try to crack the challenge!
2021-12-17 | PXD030440 | Pride
Project description:Protein N-terminal (Nt) acetylation is one of the most abundant modifications in eukaryotes, covering ~50-80 % of the proteome, depending on species. Cells with defective Nt-acetylation display a wide array of phenotypes such as impaired growth, mating defects and increased stress sensitivity. However, the pleiotropic nature of these effects has hampered our understanding of the functional impact of protein Nt-acetylation. The main enzyme responsible for Nt-acetylation throughout the eukaryotic kingdom is the N-terminal acetyltransferase NatA. Here we employed a multi-dimensional proteomics approach to analyze Saccharomyces cerevisiae lacking NatA activity, which caused global proteome remodeling. Pulsed-SILAC experiments revealed that NatA-deficient strains consistently increased degradation of ribosomal proteins compared to wild type. Explaining this phenomenon, thermal proteome profiling uncovered decreased thermostability of ribosomes in NatA-knockouts. Our data are in agreement with a role for Nt-acetylation in promoting stability for parts of the proteome by enhancing the avidity of protein-protein interactions and folding.
2023-10-24 | PXD037510 | Pride
Project description:The NF-KappaB family of transcription factors plays a critical role in numerous cellular processes, particularly the immune response. Our understanding of how the different NF-kappaB subunits act coordinately to regulate gene expression is based on a limited set of genes. We used genome-scale location analysis to identify targets of all five NF-kappaB proteins before and after stimulation of monocytic cells with bacterial lipopolysaccharide (LPS). In unstimulated cells, p50 and p52 bound to a significant number of gene promoters. p50 occupied genes together with RNA polymerase II and defined a set of genes to which other NF-kappaB proteins bound after LPS induction. In stimulated cells, genes bound by multiple NF-kappaB subunits exhibited the greatest increases in RNA polymerase II occupancy and gene expression. This study identifies novel NF-kappaB target genes, reveals how the different NF-kappaB proteins coordinate their activity and maps transcriptional regulatory networks that underlie the host response to infection.
2006-05-09 | E-WMIT-6 | biostudies-arrayexpress
Project description:Hormones and nutrients often induce genetic programs via signaling pathways that interface with gene-specific activators. Activation of the cAMP pathway, for example, stimulates cellular gene expression by means of the PKA-mediated phosphorylation of cAMP-response element binding protein (CREB) at Ser-133. Here, we use genome-wide approaches to characterize target genes that are regulated by CREB in different cellular contexts. CREB was found to occupy approximately 4,000 promoter sites in vivo, depending on the presence and methylation state of consensus cAMP response elements near the promoter. The profiles for CREB occupancy were very similar in different human tissues, and exposure to a cAMP agonist stimulated CREB phosphorylation over a majority of these sites. Only a small proportion of CREB target genes was induced by cAMP in any cell type, however, due in part to the preferential recruitment of the coactivator CREB-binding protein to those promoters. These results indicate that CREB phosphorylation alone is not a reliable predictor of target gene activation and that additional CREB regulatory partners are required for recruitment of the transcriptional apparatus to the promoter.
2006-05-09 | E-WMIT-11 | biostudies-arrayexpress