Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Proteomic analysis of integrin-associated adhesion complexes isolated from cells treated with DMSO or FAK inhibitor

ABSTRACT: Human foreskin fibroblast cells were spread on fibronectin (FN) for 1 h and treated with DMSO or FAK inhibitor (3 uM, AZ13256675 (termed AZ675 for short)) for 1 h. Integrin-associated adhesion complexes isolated from these cells were analysed by mass spectrometry to determine changes in adhesion complex composition upon FAK inhibition. As a negative control, complexes were also isolated from cells attached to transferrin (Tf), which allows integrin-independent adhesion via the transferrin receptor.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Fibroblast

SUBMITTER: Edward Horton

LAB HEAD: Martin J. Humphries

PROVIDER: PXD002720 | Pride | 2016-02-02

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	FAK dataset.pride.mztab.gz	Mztab
	FAKInhibitionTOTAL.csv	Csv
	FAKdataset.mzid.gz	Mzid
	FAKdataset.xml	Xml
	Fibronectin_DMSO_Repeat1.raw	Raw

Items per page:

1 - 5 of 15

Publications

Modulation of FAK and Src adhesion signaling occurs independently of adhesion complex composition.

Horton Edward R ER Humphries Jonathan D JD Stutchbury Ben B Jacquemet Guillaume G Ballestrem Christoph C Barry Simon T ST Humphries Martin J MJ

The Journal of cell biology 20160201 3

Integrin adhesion complexes (IACs) form mechanochemical connections between the extracellular matrix and actin cytoskeleton and mediate phenotypic responses via posttranslational modifications. Here, we investigate the modularity and robustness of the IAC network to pharmacological perturbation of the key IAC signaling components focal adhesion kinase (FAK) and Src. FAK inhibition using AZ13256675 blocked FAK(Y397) phosphorylation but did not alter IAC composition, as reported by mass spectromet ...[more]

PMID: 26833789

Similar Datasets

U2OS Adhesion Isolations +/- CytochalasinD and Neomycin

Project description:U2OS cells preferentially utilise integrin alphaV beta5 in adhesion complexes when grown under standard cell culture conditions for 3 days. AlphaV beta5 can be found in both classical 'Focal' adhesions and in novel 'Reticular' adhesions that do not associate with F-actin or other known adhesion protein components. In order to identify components of reticular adhesions, cells were treated with cytochalasinD in order to disrupt F-actin and promote loss of focal adhesions. Remaining reticular adhesion complexes were stabilised with DTBP crosslinking reagent before adhesion complexes were isolated. Preliminary data suggested that depletion of PIP2 by Neomycin treatment would lead to disruption of reticular adhesions preferentially over focal adhesions and so this manipulation was included in these experiments.

2019-06-03 | PXD008680 | Pride

HeLa and U2OS long term culture adhesion isolations

Project description:Integrin adhesion complexes have been previously isolated and analysed by mass-spectrometry from cells largely spread on matrix proteins over a couple of hours. Here we wished to determine the composition of adhesion complexes from cells cultured for 72 hours under standard cell culture conditions and identify which integrin heterodimers are utilised under these conditions.

2019-06-03 | PXD008645 | Pride

Proteomic analysis of integrin-associated adhesion complexes isolated from cells treated with DMSO or FAK inhibitor

Project description:Human foreskin fibroblast cells were spread on fibronectin (FN) for 1 h and treated with DMSO or FAK inhibitor (3 uM, AZ13256675 (termed AZ675 for short)) for 1 h. Integrin-associated adhesion complexes isolated from these cells were analysed by mass spectrometry to determine changes in adhesion complex composition upon FAK inhibition. As a negative control, complexes were also isolated from cells attached to transferrin (Tf), which allows integrin-independent adhesion via the transferrin receptor.

2016-08-23 | MSV000080089 | MassIVE

Proteomic characterisation of microtubule-dependent modulation of adhesion complexes

Project description:Investigating the effect of microtubule disruption by nocodazole treatment on isolated adhesion complexes.

2015-01-30 | PXD001183 | Pride

Integrin adhesion complex (IAC) analysis from human pancreatic ductal cells (HPDE)

Project description:Pancreatic ductal adenocarcinoma (PDAC) is characterised by an abundant desmoplastic reaction which includes an excess production of extracellular matrix and interacts with integrin adhesion receptors. Integrin adhesion complexes formed by HPDE cells (H6c7), with and without expression of mutant KRas G12V, were isolated and analysed by LC-MSMS

2022-05-20 | PXD027827 | Pride

Proteomic analysis of integrin adhesion complex disassembly induced by nocodazole

Project description:To explore the dynamics of integrin adhesion complex disassembly, isolated adhesion complexes from U2OS cells were subjected to mass spectrometry based proteomic analysis. U2OS cells were allowed to spread on FN, treated with nocodazole for 4 hours to disrupt microtubules. To coordinate microtubule-induced integrin adhesion complex disassembly nocodazole was subsequently washed out and adhesion complexes isolated at 5, 10 and 15 minutes during this process.

2015-10-14 | PXD002129 | Pride

Mesenchymal stem cell integrin-associated complexes

Project description:The composition and stiffness of the extracellular matrix (ECM) environment that surrounds Multipotent mesenchymal stem cells (MSCs) stem cells dictates transcriptional programming, thereby affecting stem cell lineage decision-making. Cells sense force between themselves and their microenvironment controlling the capability of MSCs to differentiate into adipocytes, osteocytes and chondrocytes. Force sensing is transmitted by integrin receptors and their associated adhesion signalling complexes. To identify regulators of MSC force sensing we sought to catalogue MSC adhesion complex composition. Therefore we isolated integrin-associated adhesion complexes formed in MSCs plated on the ECM ligand fibronectin. We identifed proteins using mass spectrometry that define a MSC specific subset of adhesion complex proteins consisting of key linkages to the actin cytoskeleton together with integrin signalling and force sensing components.

2015-07-20 | PXD001809 | Pride

Proteomic analysis of integrin adhesion complex assembly

Project description:To explore the dynamics of integrin adhesion complex assembly, isolated adhesion complexes from K562 cells were subjected to mass spectrometry based proteomic analysis. K562 cells were incubated with FN-coated beads for 1, 7 and 30 minutes and protein complexes stabilised for 2 minutes with DTBP crosslinking reagent before adhesion complexes were isolated.

2015-10-14 | PXD002159 | Pride

The Itgb3 Adhesome - Integrin β3 is seen as a key anti-angiogenic target for cancer treatment due to its expression on neovasculature

Project description:Integrin β3 is seen as a key anti-angiogenic target for cancer treatment due to its expression on neovasculature, but the role it plays in the process is complex; whether it is pro- or anti-angiogenic depends on the context in which it is expressed. To understand precisely β3’s role in regulating integrin adhesion complexes in endothelial cells, we characterised, by mass spectrometry, the β3-dependent adhesome. We show that depletion of β3-integrin in this cell type leads to changes in microtubule behaviour that control their migration. β3-integrin regulates microtubule stability in endothelial cells through Rcc2/Anxa2 driven control of active Rac1 localisation. Our findings reveal that angiogenic processes, both in vitro and in vivo, are more sensitive to microtubule targeting agents when β3-integrin levels are reduced.

2018-05-23 | PXD008591 | Pride

Expression data from FAK null mouse embryonic fibroblasts treated with endothelin-1

Project description:Endothelin-1 (ET-1) plays a critical role in connective tissue remodeling by fibroblasts during tissue repair and fibrosis. We investigated the molecular pathways in the transmission of ET-1 signals that lead to features of connective tissue remodeling, in particular the role of FAK (focal adhesion kinase). We used microarrays to investigated whether FAK is required for ET-1 to promote the global programme of gene expression underlying myofibroblast formation and identified distinct classes of up-regulated genes during this process. Genes whose induction by ET-1 required FAK Affymetrix Mouse Genome 430 2.0 GeneChips were used to identify differential gene expression changes bewteen samples.

2012-06-30 | E-GEOD-30356 | biostudies-arrayexpress