Browse
Submit Data
Databases
API
Help

Dataset Information

175 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

A selective photoaffinity probe enables assessment of cannabinoid CB2 receptor expression and ligand engagement in human cells

ABSTRACT: Chemical tools and methods that report on G protein-coupled receptor (GPCR) expression levels and receptor occupancy by small molecules are highly desirable. We report the development of LEI121 as a photoreactive probe to study the type 2 cannabinoid receptor (CB2R), a promising GPCR to treat tissue injury and inflammatory diseases. LEI121 is the first CB2R-selective bifunctional probe that covalently captures CB2R upon photoactivation. An incorporated alkyne serves as ligation handle for the introduction of reporter groups. LEI121 enables target engagement studies and visualization of endogenously expressed CB2R in HL-60 as well as primary human immune cells using flow cytometry. Our findings show that strategically functionalized probes allow monitoring of endogenous GPCR expression and engagement in human cells using tandem photo-click chemistry and hold promise as biomarkers in translational drug discovery.

INSTRUMENT(S): LTQ Orbitrap, Synapt MS

ORGANISM(S): Homo Sapiens (human) Cricetulus Griseus (chinese Hamster) (cricetulus Barabensis Griseus)

TISSUE(S): Epithelial Cell, Cell Culture

DISEASE(S): Disease Free

SUBMITTER: Bogdan Florea

LAB HEAD: Mario van der Stelt

PROVIDER: PXD008460 | Pride | 2018-06-15

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	F015250.xml	Xml
	F015251.xml	Xml
	F015252.xml	Xml
	F015255.xml	Xml
	TableS2-40kDaregion.zip.raw	Raw

Items per page:

1 - 5 of 42

Similar Datasets

photoaffinity for Puerarin and mouse brainstem

Project description:To elucidate the molecular mechanism mediating the inactivated effect of DMV neurons on fat absorption, we performed an activity-based protein profiling strategy, using Puerarin as a “bait”. The Puerarin-tag probe was synthesized with a photoreactive tag to enrich and visualize target proteins via a photoaffinity chemistry reaction. We verified that probe-tagged Puerarin retains the same effects of increasing fecal lipid excretion as non-tagged Puerarin. Probe-tagged Puerarin was added to the freshly isolated brainstem sample, and 10 doses of non-tagged Puerarin was used as a competitor of probe-tagged Puerarin. Following the photoaffinity reaction, targeted proteins were subsequently assessed by liquid chromatography tandem mass spectrometry (LC-MS).

2024-09-06 | PXD052140 | Pride

RNA sequencing to determine the contribution of kinase receptor transactivation to G protein coupled receptor signalling in vascular smooth muscle cells

Project description:G protein coupled receptor (GPCR) signalling covers three major mechanisms. GPCR agonist engagement allows for the G proteins to bind to the receptor leading to a classical downstream signalling cascade. The second mechanism is via the utilization of the β-arrestin signalling molecule and thirdly via transactivation dependent signalling. GPCRs can transactivate protein tyrosine kinase receptors (PTKR) to activate respective downstream signalling intermediates. In the past decade GPCR transactivation dependent signalling was expanded to show transactivation of serine/threonine kinase receptors (S/TKR). Kinase receptor transactivation enormously broadens the GPCR signalling paradigm. This work utilizes next generation RNA-sequencing to study the contribution of transactivation dependent signalling to total protease activated receptor (PAR)-1 signalling. Transactivation, assessed as gene expression, accounted for 50 percent of the total genes regulated by thrombin acting through PAR-1 in human coronary artery smooth muscle cells. GPCR transactivation of PTKRs is approximately equally important as the transactivation of the S/TKR with 209 and 177 genes regulated respectively, via either signalling pathway. This work shows that genome wide studies can provide powerful insights into GPCR mediated signalling pathways

2017-09-05 | GSE99770 | GEO

Selectivity profiling of SLC16A3/MCT4 inhibitor slCeMM1 using photoaffinity probe

Project description:To assess the selectivity of SLC16A3/MCT4 inhibitor slCeMM1, we derived diazirine-alkyne photoaffinity probe (slCeMM1-PAP). We next used HAP1 and MDAMB231 cell lines for MS experiment, comparing the enrichment of proteins by slCeMM1-PAP and conditions where slCeMM1-PAP was competed with slCeMM1 or it’s structurally related inactive control (S1_007). We found that among proteins enriched by slCeMM1-PAP, only SLC16A3 was competed by slCeMM1, but not S1_007, confirming the SLC16A3 engagement by slCeMM1 and suggesting overall good selectivity of slCeMM1.

2023-10-16 | PXD040089 | Pride

Engagement of TCR with antibodies

Project description:Comparison of gene expression changes in CD4+CD8+ thymocytes following engagement of TCR with anti-Valpha or Vbeta antibodies Keywords: parallel sample

2005-01-01 | GSE1448 | GEO

An amygdala circuit that supresses social engagement

Project description:An amygdala circuit that supresses social engagement

| PRJNA703639 | ENA

Transcription profiling of mouse CD4+CD8+ thymocytes following engagement of TCR with anti-Valpha or Vbeta antibodies

Project description:Comparison of gene expression changes in CD4+CD8+ thymocytes following engagement of TCR with anti-Valpha or Vbeta antibodies

2007-09-28 | E-GEOD-1448 | biostudies-arrayexpress

Mapping in vivo target interaction profiles of covalent inhibitors using chemical proteomics with label-free quantification

Project description:Activity-based protein profiling (ABPP) has emerged as a valuable chemical proteomics method to guide the therapeutic development of covalent drugs by assessing their on-target engagement and off-target activity. Recently, we reported on the establishment, by ABPP, of the serine hydrolase interaction landscape of the experimental drug BIA 10-2474, thereby providing a potential explanation for the adverse side effects observed with this compound. ABPP allows mapping of protein interaction landscapes of inhibitors in cells, tissues and animal models. Here, we describe a protocol for identifying the in vivo selectivity profile of covalent inhibitors using ABPP with label-free quantitative proteomics as exemplified by experimental data on the diacylglycerol lipase inhibitor DH376 in mouse brain, liver, kidney and testes. The protocol has been optimized using label-free quantification methods to have high proteome coverage and to allow the simultaneous comparison of multiple biological samples. The stages of the protocol include tissue lysis, probe incubation, enrichment, sample preparation, liquid chromatography-mass spectrometry measurement, processing and data analysis. The results include evidence of target engagement in a native proteome and the identification of potential off-targets for the inhibitor under investigation. The entire protocol takes at least four days (depending on the number of samples).

2018-02-23 | PXD007965 | Pride

Tracking drug action on the cell surface proteome

Project description:Endogenous ligands and drugs interact with the cell-surface proteome but hypothesis-free tracking of dynamic remodeling processes at the plasma membrane is challenging. We introduce cell surface thermal proteome profiling for comprehensive characterization of ligand-induced changes in protein abundances and thermal stabilities. We demonstrate drug binding to extracellular receptors, complexes and transporters, discover stimulation-dependent remodeling of T-cell receptor complexes and describe a competition-based approach to measure target engagement of GPCR antagonists.

2020-10-26 | PXD016249 | Pride

Dichotomous Engagement of HDAC3 Activity Governs Inflammatory Responses

Project description:Dichotomous Engagement of HDAC3 Activity Governs Inflammatory Responses

| PRJNA590241 | ENA

Affinity and Dose of TCR Engagement Yield Proportional Enhancer and Gene Activity in CD4+ T Cells

Project description:Affinity and dose of T cell receptor (TCR) interaction with antigens govern the magnitude of CD4+ T cell responses, but questions remain regarding the quantitative translation of TCR engagement into downstream signals. We find that while the response of CD4+ T cells to antigenic stimulation is bimodal, activated cells exhibit analog responses proportional to signal strength. Gene expression output reflects TCR signal strength, providing a signature of T cell activation. Expression changes rely on a pre-established enhancer landscape and quantitative acetylation at AP-1 binding sites. Finally, we show that graded expression of activation genes depends on ERK pathway activation, suggesting that an ERK-AP-1 axis translates TCR signal strength into proportional activation of enhancers and genes essential for T cell function. CD4+ T cells from transgenic AND mice were sequenced under the conditions indicated. Replicates are included for each type of data (RNA-Seq, ChIP-Seq), and are numbered accordingly. The No Peptide condition serves as the untreated control for the peptide-treated samples, and inputs are provided for ChIP-Sequencing samples.

2016-06-01 | E-GEOD-69545 | biostudies-arrayexpress

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