Mass spectrometry is a powerful tool for identification of proteins associated with lipid rafts of Jurkat T-cell line
Language English Country Great Britain, England Media print
Document type Journal Article, Research Support, Non-U.S. Gov't, Review
PubMed
15494013
DOI
10.1042/bst0320777
PII: BST0320777
Knihovny.cz E-resources
- MeSH
- Cell Adhesion MeSH
- Cell Membrane metabolism MeSH
- Centrifugation, Density Gradient MeSH
- Cytoskeleton metabolism MeSH
- Detergents pharmacology MeSH
- Spectrometry, Mass, Electrospray Ionization MeSH
- Mass Spectrometry methods MeSH
- Ions MeSH
- Jurkat Cells MeSH
- Humans MeSH
- Membrane Microdomains metabolism MeSH
- Proteins chemistry MeSH
- Sucrose pharmacology MeSH
- Signal Transduction MeSH
- Statistics as Topic methods MeSH
- Protein Binding MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Review MeSH
- Names of Substances
- Detergents MeSH
- Ions MeSH
- Proteins MeSH
- Sucrose MeSH
Many proteins involved in signal-transduction pathways are concentrated in membrane microdomains enriched in lipids with distinct physical properties. Since these microdomains are insoluble in non-ionic detergents in cold, proteins associated with them could be efficiently purified by techniques such as sucrose-density gradient centrifugation. The complexity of the resulting protein mixture requires powerful MS technique for its analysis. We have found that successful identification of biologically relevant proteins is critically dependent on the enrichment of the starting material (plasma membranes), and on the extraction procedure. Applying these conditions in combination with microHPLC-ESI (electrospray ionization)-MS/MS, we have identified proteins involved in signalling, cytoskeletal association and cellular adhesion in Jurkat cells that are not stimulated by any antibody incubation.
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