Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

A chemoproteomic platform for selective deubiquitinase inhibitor discovery

ABSTRACT: Here we describe our work on the development of an ABPP screening platform for cysteine reactive covalent fragments against deubiquitinating enzymes (DUBs) on 96 well plate format. Within this targeted workflow, we profile cysteine reactive fragments by competition with biotinylated ubiquitin probe in cell lysates and employ label free data independent acquisition (DIA) MS method. The platform was used to screen a library of 138 Cys-reactive covalent fragments against DUBome in HEK293T cell lysate, identifying functionally relevant hit fragments for numerous DUBs and demonstrating the utility of the approach in expanding the liganded proteome.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Aini Vuorinen

LAB HEAD: Jacob Bush

PROVIDER: PXD037152 | Pride | 2024-03-18

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	200605_DC1927_HEK_bead_a_1.raw	Raw
	200605_DC1927_HEK_bead_a_2.raw	Raw
	200605_DC1927_HEK_bead_b_1.raw	Raw
	200605_DC1927_HEK_bead_b_2.raw	Raw
	200605_DC1927_PA_0pt5_cmp_a_1.raw	Raw

Items per page:

1 - 5 of 260

Similar Datasets

A chemoproteomic platform for selective deubiquitinase inhibitor discovery

Project description:Here we describe our work on the development of an ABPP screening platform for cysteine reactive covalent fragments against deubiquitinating enzymes (DUBs) on 96 well plate format. Within this targeted workflow, we profile cysteine reactive fragments by competition with biotinylated ubiquitin probe in cell lysates and employ label free data independent acquisition (DIA) MS method. The platform was used to screen a library of 138 Cys-reactive covalent fragments against DUBome in HEK293T cell lysate, identifying functionally relevant hit fragments for numerous DUBs and demonstrating the utility of the approach in expanding the liganded proteome.

2024-03-18 | PXD037114 | Pride

Profiling Sulfur(VI) Fluorides as Reactive Functionalities for Chemical Biology Tools and Expansion of the Ligandable Proteome

Project description:Here, we profiled sulfur(VI) fluorides (SVI-F) as reactive functionalities that can modify multiple residues including Lys, Tyr, His and Ser, thus offering utility for targeting almost any protein. These studies provided an in-depth understanding of SVI-F functionalities, including hydrolytic stability, protein reactivity and utility in chemoproteomics

2024-03-18 | PXD037048 | Pride

The Proteomic Landscape of in vitro Cultured Endothelial Cells across Vascular Beds

Project description:Blood vessel endothelial cells (EC) display heterogeneity across vascular beds, which is at the bases of organ specificity and anticipated to drive site-specific vascular pathology. It is assessed in vivo using transcriptomics, and in vitro using functional assays, but how proteomes compare across human in vitro cultured ECs remains incompletely characterized. We generated an in-depth human EC proteomic landscape (>8000 proteins) across 6 vascular beds and 2 in vitro models both in steady-state and upon IFNγ-induced inflammation. EC proteomes displayed a high similarity and organ specific proteins were limited. Variation between ECs and donors was mainly based in biological processes underlying proliferation and differentiation. BOECs and HUVECs represented the extremes of proteomic phenotypes. IFNγ responses were highly conserved across cellular sources. Harnessing dynamics in protein abundances we delineated interactor networks of VWF and VE-Cadherin. This EC landscape provides an extensive proteomic addition in studying EC biology and heterogeneity from an in vitro perspective.

2024-07-25 | PXD045899 | Pride

Metabolic crosstalk between skeletal muscle cells and liver through IRF4-FSTL1 in 2 nonalcoholic steatohepatitis

Project description:In this study Guo et al. revealed an endocrine pathway regulated by skeletal muscle IRF4 that manipulates liver pathology. Mice with skeletal muscle specific ablation of IRF4 show ameliorated liver steatosis, inflammation, and fibrosis, without changes in body weight on nonalcoholic steatohepatitis (NASH) diet. Proteomics analysis of mouse serum suggested that follistatin-like protein 1 (FSTL1) might link the communication between muscle and liver. Dual luciferase assays showed that IRF4 could transcriptionally regulate FSTL1 and reconstitution of FSTL1 expression in muscle of F4MKO mice was sufficient to restore the liver pathology. Furthermore, co-culture experiments verified that FSTL1 exerts its function in hepatocyte, macrophage, and hepatic satellite cells through CD14 and DIP2A, CD14, DIP2A, respectively. In human, serum FSTL1 is increased in NASH patients, but the mRNA level of Fstl1 and its receptors are decreased in NASH liver biopsy, suggesting the increased serum FSTL1 is from skeletal muscle as studied in F4MKO mice. These data unveiled a signaling pathway from skeletal muscle to liver via IRF4-FSTL1-DIP2A/CD14 in the pathogenesis of NASH.

2023-09-13 | PXD045324 | Pride

Species-specific metabolic reprogramming in human and mouse microglia during inflammatory pathway induction

Project description:Metabolic reprogramming is a hallmark of the immune cells in response to inflammatory stimuli. This metabolic process involves a switch from oxidative phosphorylation (OXPHOS)to glycolysis, or alterations in other metabolic pathways. However, most of the experimental findings have been acquired in murine immune cells and little is known about the metabolic reprogramming of human microglia. In this study, we investigated the transcriptomic and metabolic profiles of mouse and iPSC-derived human microglia challenged with the TLR4 agonist LPS. We found that both species displayed a metabolic shift and an overall increased glycolytic gene signature in response to LPS treatment. The metabolic reprogramming was characterized by the upregulation of hexokinases in mouse microglia and phosphofructokinases in human microglia. This study provides the first direct comparison of energy metabolism between mouse and human microglia, highlighting the species-specific pathways involved in immunometabolism and the importance of considering these differences in translational research.

2023-09-02 | PXD043836 | Pride

SDH deficiency-driven succinate accumulation dictates the drug resistance of AML through regulating p-UBC12/cullin activity

Project description:Drug resistance is vital for the poor prognosis of acute myeloid leukemia (AML) patients, but the underlying mechanism remains poorly understood. Given the unique microenvironment of bone marrow, we reasoned that drug resistance of AML might rely on distinct microenvironment-associated metabolic processes. Here, we identified SDH deficiency and over-cumulative succinate as typical features in AML, with a marked function in causing the resistance of AML cells to a wide range of anti-cancer therapies. Mechanistically, succinate promoted the accumulation of oncogenic proteins in a manner that precedes transcriptional activation. This function was mediated by succinate-triggered upregulation of UBC12 phosphorylation, which impaired its E2 function in cullins neddylation. Notably, decreasing succinate levels by fludarabine could effectively restore the drug sensitivity of SDH-deficient AML PDX. Together, we uncover a novel function of succinate in driving drug resistance by regulating p-UBC12/cullin activity, and indicate reshaping succinate metabolism as a promising treatment for SDH-deficient AML.

2024-08-28 | PXD047175 | Pride

Methylmalonic acidemia triggers lysosomal-autophagy dysfunctions

Project description:Methylmalonic acidemia (MMA) is a rare inborn error of propionate metabolism caused by deficiency of the mitochondrial methylmalonyl-CoA mutase (MUT) enzyme. As matter of fact, MMA patients manifest impairment of the primary metabolic network with profound damages that involve several cell components, many of which have not been discovered yet. We employed cellular models and patients-derived fibroblasts to refine and uncover new pathologic mechanisms connected with MUT deficiency through the combination of multi-proteomics and bioinformatics approaches.

2024-05-23 | PXD044101 | Pride

High quality MS/MS spectrum prediction for data-dependent and -independent acquisition data analysis

Project description:Peptide fragmentation spectra are routinely predicted in the interpretation of mass spectrometry-based proteomics data. Unfortunately, the generation of fragment ions is not well enough understood to estimate fragment ion intensities accurately. Here, we demonstrate that machine learning can predict peptide fragmentation patterns in mass spectrometers with accuracy within the uncertainty of the measurements. Moreover, analysis of our models reveals that peptide fragmentation depends on long-range interactions within a peptide sequence. We illustrate the utility of our models by applying them to the analysis of both data-dependent and data-independent acquisition datasets. In the former case, we observe a significant increase in the total number of peptide identifications at fixed false discovery rate. In the latter case we demonstrate that the use of predicted MS/MS spectra is equivalent to the use of spectra from experimentallibraries, indicating that fragmentation libraries for proteomics are becoming obsolete.

2019-03-13 | PXD010382 | Pride

Proteomic changes to immune and inflammatory processes underlie lung preservation using ex vivo cytokine adsorption

Project description:The field of graft preservation has made considerable strides in recent years improving outcomes related to solid organ restoration and regeneration. In lungs, the use of ex vivo lung perfusion (EVLP) in line with devices and treatments has shown promising results within preclinical and clinical studies with the potential to improve graft quality. The benefit of the therapy would be to render marginal and declined donor lungs suitable for transplantation, ultimately increasing the donor pool available for transplantation. Additionally, such therapies used in machine perfusion could also increase preservation time, facilitating logistical planning. Cytokine adsorption has been demonstrated as a potentially safe and effective therapy when applied to the EVLP circuit and post transplantation. The mechanism by which this treatment improves the donor lung on a molecular basis is not yet fully elucidated. We hypothesized that there were characteristic inflammatory and immunomodulatory differences between lungs treated with and without cytokine adsorption, reflecting in proteomic changes in gene ontology pathways and across inflammation-related proteins. In the current study we investigate the molecular mechanisms and signaling pathways of how cytokine adsorption impacts the lung function when used during EVLP and when used post transplantation as hemoperfusion in a porcine model. Lung tissue from EVLP and post lung transplantation were analyzed for their proteomic profile using mass spectrometry. The inflammatory and immune processes were compared between the treated and the non-treated groups to show the differences occurring between the forms of graft preservation.

2023-10-24 | PXD044413 | Pride

Guanosine-7 tRNA methylation steers tRNA-derived fragment biogenesis and translational control in prostate cancer

Project description:Transfer RNAs (tRNAs) are exceptionally subject to modifications, including methylation. While mRNA methylation is emerging as an important regulator of biological and pathological processes in cancer, how post-transcriptional methylation of tRNAs contributes to cancer is largely unknown. Here we show that the RNA N7-methylguanosine (m7G) methyltransferase METTL1 is highly differentially expressed in prostate cancer compared to non-tumour prostate tissues. METTL1 expression regulation is mediated under the oncogenic PI3K-PTEN pathway. Knockdown of METTL1 dramatically inhibits prostate cancer cell growth and tumour progression in vivo. In contrast, overexpression of the wild type but not the catalytically inactive METTL1 potentiates cell growth. Thus, METTL1-mediated methylation is important for prostate tumorigenesis. Mechanistically we find that METTL1 depletion causes loss of m7G tRNA methylation and increases endonucleolytic cleavage of tRNA leading to an accumulation of 5′ tRNA-derived small RNA fragments. 5′ tRNA-derived fragments steer translation control to favour synthesis of key regulators of tumour growth suppression and immune rejection. In summary, our findings uncover a critical function of m7G tRNA methylation in directing translation control in cancer cells with important implications for tumour growth and unveil METTL1 inhibition as a promising anti-cancer therapeutic strategy.

2023-10-24 | PXD030141 | Pride

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data