Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

HeLa and U2OS long term culture adhesion isolations

ABSTRACT: 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.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell

DISEASE(S): Cervical Cancer,Osteosarcoma

SUBMITTER: Matthew Jones

LAB HEAD: Martin J Humphries

PROVIDER: PXD008645 | Pride | 2019-06-03

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Publications

Reticular adhesions are a distinct class of cell-matrix adhesions that mediate attachment during mitosis.

Lock John G JG Jones Matthew C MC Askari Janet A JA Gong Xiaowei X Oddone Anna A Olofsson Helene H Göransson Sara S Lakadamyali Melike M Humphries Martin J MJ Strömblad Staffan S

Nature cell biology 20181022 11

Adhesion to the extracellular matrix persists during mitosis in most cell types. However, while classical adhesion complexes, such as focal adhesions, do and must disassemble to enable mitotic rounding, the mechanisms of residual mitotic cell-extracellular matrix adhesion remain undefined. Here, we identify 'reticular adhesions', a class of adhesion complex that is mediated by integrin αvβ5, formed during interphase, and preserved at cell-extracellular matrix attachment sites throughout cell div ...[more]

PMID: 30361699

Similar Datasets

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.

2016-02-02 | PXD002720 | Pride

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

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

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

The tongue squamous carcinoma cell line Cal27 primarily employs integrin α6β4-containing type II hemidesmosomes for adhesion which contribute to anticancer drug sensitivity

Project description:Integrins are heterodimeric cell surface glycoproteins used by cells to bind to the extracellular matrix (ECM) and regulate tumor cell proliferation, migration and survival. A causative relationship between integrin expression and resistance to anticancer drugs has been demonstrated in different tumors, including head and neck squamous cell carcinoma. Using a Cal27 tongue squamous cell carcinoma model, we have previously demonstrated that de novo expression of integrin αVβ3 confers resistance to several anticancer drugs (cisplatin, mitomycin C and doxorubicin) through a mechanism involving downregulation of active Src, increased cell migration and invasion. In the integrin αVβ3 expressing Cal27-derived cell clone 2B1, αVβ5 expression was also increased, but unrelated to drug resistance. To identify the integrin adhesion complex (IAC) components that contribute to the changes in Cal27 and 2B1 cell adhesion and anticancer drug resistance, we isolated IACs from both cell lines. Mass spectrometry (MS)-based proteomics analysis indicated that both cell lines preferentially use integrin α6β4, which is classically found in hemidesmosomes. The anticancer drug resistant cell clone 2B1 demonstrated an increased level of α6β4 accompanied with increased deposition of a laminin-332-containing ECM. Immunofluorescence and electron microscopy demonstrated the formation of type II hemidesmosomes by both cell types. Furthermore, suppression of α6β4 expression in both lines conferred resistance to anticancer drugs. Taken together, our results identify a key role for α6β4-containing type II hemidesmosomes in regulating anticancer drug sensitivity.

2022-02-16 | PXD028810 | 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

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 melanoma cell line MDA-MB-435 adhesome in long-term culture and its comparison to the adhesome upon integrin αV knockdown

Project description:Using mass spectrometry–based proteomics, we analysed the components of integrin adhesion complexes of MDA-MB-435S cells and two MDA-MB-435S-derived integrin αV-specific shRNA-expressing cell clones. Among integrins, we detected αV and β5 integrin subunits in the MDA-MB-435S adhesome, thus showing that in long term culture these cells use preferentially integrin αVβ5 for adhesion. Our data represents the first published adhesome of αVβ5 integrin heterodimer.

2020-02-17 | PXD016837 | 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

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

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