Proteomics

Dataset Information

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Membrane enriched proteome of transfected SW480 cells


ABSTRACT: This project used membrane-enriched proteomics to compare the membrane proteome of SW480 human colorectal cancer subclones that had been transfected with the full length coding sequence of the beta-6 integrin subunit under an overexpression promoter (SW480OE) against SW480 cells that had been transfected with an ‘empty’ mock vector (SW480M). Cell lysate samples were enriched for membrane proteins using a modified sodium carbonate stripping method before undergoing peptide immobilised pH gradient isoelectric focusing (IPG-IEF). Two IPG-IEF pH ranges were used to increase proteomic coverage: pH3-10 (broad range; BR) and pH3.5-4.5 (narrow range; NR). Each IPG-IEF strip was cut into 24 equal fractions and the peptides eluted. IPG-IEF of each pH range for both cell lines was performed in triplicate, resulting in four triplicate sets of 24 fractions. Data processing and bioinformatics: Spectra were identified against the *Homo sapiens* protein database (derived from Ensembl, Swiss-Prot and NCBI) using X!Tandem and the Global Proteome Machine (GPM) Tornado XE (build date 2010.12.01). GPM searches were set to tryptic peptides under GPM’s default fragment ion mass tolerance of 0.4Da and did not include semi-cleavage. Searches included single acid polymorphisms and default GPM residue modifications (oxidation,dioxidation, carbamidomethyl, deamidation, acetylation).

INSTRUMENT(S): LTQ

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: David Cantor  

PROVIDER: PXD000230 | Pride | 2013-05-30

REPOSITORIES: Pride

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Publications

Overexpression of αvβ6 integrin alters the colorectal cancer cell proteome in favor of elevated proliferation and a switching in cellular adhesion that increases invasion.

Cantor David D   Slapetova Iveta I   Kan Alison A   McQuade Leon R LR   Baker Mark S MS  

Journal of proteome research 20130524 6


Many proteins enhance cancer progression toward life-threatening metastases. These include linking proteins called integrins that mediate cell adhesion to the extracellular matrix (ECM), consequently altering both function and phenotype. Specific neoexpression of the β6 integrin subunit correlates with the epithelial-to-mesenchymal transition, metastasis, and poor overall patient survival. While β6 is implicated in these processes, exactly how it affects signaling and/or proteolytic pathways in  ...[more]