Proteomics

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Global Membrane Protein Interactome Analysis using In vivo Crosslinking and MS-based Protein Correlation Profiling

ABSTRACT: We present a methodology using in vivo crosslinking combined with HPLC-MS for the global analysis of endogenous protein complexes by protein correlation profiling. Formaldehyde crosslinked protein complexes were extracted with high yield using denaturing buffers that maintained complex solubility during chromatographic separation. We show this efficiently detects both integral membrane and membrane-associated protein complexes, in addition to soluble complexes, allowing identification and analysis of complexes not accessible in native extracts. We compare the protein complexes detected by HPLC-MS protein correlation profiling in both native and formaldehyde crosslinked U2OS cell extracts. These proteome-wide datasets of both in vivo crosslinked and native protein complexes from U2OS cells are freely available via a searchable online database (www.peptracker.com/epd).

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Dr Angus Lamond 

PROVIDER: MSV000080014 | MassIVE | Tue Aug 02 16:24:00 BST 2016

SECONDARY ACCESSION(S): PXD003754

REPOSITORIES: MassIVE

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Global Membrane Protein Interactome Analysis using In vivo Crosslinking and Mass Spectrometry-based Protein Correlation Profiling.

Larance Mark M   Kirkwood Kathryn J KJ   Tinti Michele M   Brenes Murillo Alejandro A   Ferguson Michael A J MA   Lamond Angus I AI  

Molecular & cellular proteomics : MCP 20160425 7

We present a methodology using in vivo crosslinking combined with HPLC-MS for the global analysis of endogenous protein complexes by protein correlation profiling. Formaldehyde crosslinked protein complexes were extracted with high yield using denaturing buffers that maintained complex solubility during chromatographic separation. We show this efficiently detects both integral membrane and membrane-associated protein complexes,in addition to soluble complexes, allowing identification and analysi  ...[more]

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