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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November 2004
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Conference Article|
October 26 2004
Mass spectrometry is a powerful tool for identification of proteins associated with lipid rafts of Jurkat T-cell line
P. Pompach;
P. Pompach
1
*Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic
†Department of Biochemistry, Faculty of Science, Charles University, Hlavova 8, CZ-12840 Prague, Czech Republic
1To whom correspondence should be addressed (email pompach@biomed.cas.cz).
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P. Man;
P. Man
*Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic
†Department of Biochemistry, Faculty of Science, Charles University, Hlavova 8, CZ-12840 Prague, Czech Republic
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P. Novák;
P. Novák
*Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic
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V. Havlíček;
V. Havlíček
*Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic
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A. Fišerová;
A. Fišerová
*Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic
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K. Bezouška
K. Bezouška
*Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic
†Department of Biochemistry, Faculty of Science, Charles University, Hlavova 8, CZ-12840 Prague, Czech Republic
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Publisher: Portland Press Ltd
Received:
July 19 2004
Online ISSN: 1470-8752
Print ISSN: 0300-5127
© 2004 The Biochemical Society
2004
Biochem Soc Trans (2004) 32 (5): 777–779.
Article history
Received:
July 19 2004
Citation
P. Pompach, P. Man, P. Novák, V. Havlíček, A. Fišerová, K. Bezouška; Mass spectrometry is a powerful tool for identification of proteins associated with lipid rafts of Jurkat T-cell line. Biochem Soc Trans 1 November 2004; 32 (5): 777–779. doi: https://doi.org/10.1042/BST0320777
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