Projects per year
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). Raw data are also available via ProteomeXchange (identifier PXD003754).
Original language | English |
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Pages (from-to) | 2476-2490 |
Number of pages | 15 |
Journal | Molecular & Cellular Proteomics |
Volume | 15 |
Issue number | 7 |
Early online date | 25 Apr 2016 |
DOIs | |
Publication status | Published - 1 Jul 2016 |
Keywords
- membrane protein
- interactome analysis
- proteomics
- protein complexes
- protein cross-linking
- mass spectrometry (MS)
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Dive into the research topics of 'Global membrane protein interactome analysis using in vivo crosslinking and MS-based protein correlation profiling'. Together they form a unique fingerprint.Projects
- 2 Finished
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Multidimensional Proteomic Analysis of Metabolic Stress & Cellular Phenotypes (Strategic Grant)
Cantrell, D. (Investigator) & Lamond, A. (Investigator)
1/01/15 → 31/12/19
Project: Research
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Protein Glycosylation in Trypanosomes: Defining and Exploiting a Biological System (Senior Investigator Award)
Ferguson, M. (Investigator)
1/10/13 → 30/06/23
Project: Research