Proteomic analysis of malignant lymphocyte membrane microparticles using double ionization coverage optimization
Laurent Miguet
Laboratoire de Spectrométrie de Masse Bio-Organique, UMR 7512, Strasbourg, France
Search for more papers by this authorKarine Pacaud
Laboratoire de Spectrométrie de Masse Bio-Organique, UMR 7512, Strasbourg, France
Search for more papers by this authorClaire Felden
Institut d'Hématologie et d'Immunologie, Faculté de Médecine, Université Louis Pasteur, Strasbourg Cedex, France
Search for more papers by this authorBénédicte Hugel
Institut d'Hématologie et d'Immunologie, Faculté de Médecine, Université Louis Pasteur, Strasbourg Cedex, France
Search for more papers by this authorM. Carmen Martinez
Institut d'Hématologie et d'Immunologie, Faculté de Médecine, Université Louis Pasteur, Strasbourg Cedex, France
Search for more papers by this authorJean-Marie Freyssinet
Institut d'Hématologie et d'Immunologie, Faculté de Médecine, Université Louis Pasteur, Strasbourg Cedex, France
Search for more papers by this authorRaoul Herbrecht
Département d'Onco-Hématologie, Hôpital Hautepierre, Strasbourg, France
Search for more papers by this authorCorresponding Author
Noelle Potier Dr.
Laboratoire de Spectrométrie de Masse Bio-Organique, UMR 7512, Strasbourg, France
Laboratoire de Spectrométrie de Masse Bio-Organique, ECPM, Université Louis Pasteur, CNRS UMR 7512, 25 rue Becquerel 67087 Strasbourg Cedex 2 France Fax: +33-390-242781===Search for more papers by this authorAlain van Dorsselaer
Laboratoire de Spectrométrie de Masse Bio-Organique, UMR 7512, Strasbourg, France
Search for more papers by this authorLaurent Mauvieux Dr.
Institut d'Hématologie et d'Immunologie, Faculté de Médecine, Université Louis Pasteur, Strasbourg Cedex, France
Laboratoire d'Hématologie, Hôpital Hautepierre, Strasbourg, France
Additional corresponding author
Search for more papers by this authorLaurent Miguet
Laboratoire de Spectrométrie de Masse Bio-Organique, UMR 7512, Strasbourg, France
Search for more papers by this authorKarine Pacaud
Laboratoire de Spectrométrie de Masse Bio-Organique, UMR 7512, Strasbourg, France
Search for more papers by this authorClaire Felden
Institut d'Hématologie et d'Immunologie, Faculté de Médecine, Université Louis Pasteur, Strasbourg Cedex, France
Search for more papers by this authorBénédicte Hugel
Institut d'Hématologie et d'Immunologie, Faculté de Médecine, Université Louis Pasteur, Strasbourg Cedex, France
Search for more papers by this authorM. Carmen Martinez
Institut d'Hématologie et d'Immunologie, Faculté de Médecine, Université Louis Pasteur, Strasbourg Cedex, France
Search for more papers by this authorJean-Marie Freyssinet
Institut d'Hématologie et d'Immunologie, Faculté de Médecine, Université Louis Pasteur, Strasbourg Cedex, France
Search for more papers by this authorRaoul Herbrecht
Département d'Onco-Hématologie, Hôpital Hautepierre, Strasbourg, France
Search for more papers by this authorCorresponding Author
Noelle Potier Dr.
Laboratoire de Spectrométrie de Masse Bio-Organique, UMR 7512, Strasbourg, France
Laboratoire de Spectrométrie de Masse Bio-Organique, ECPM, Université Louis Pasteur, CNRS UMR 7512, 25 rue Becquerel 67087 Strasbourg Cedex 2 France Fax: +33-390-242781===Search for more papers by this authorAlain van Dorsselaer
Laboratoire de Spectrométrie de Masse Bio-Organique, UMR 7512, Strasbourg, France
Search for more papers by this authorLaurent Mauvieux Dr.
Institut d'Hématologie et d'Immunologie, Faculté de Médecine, Université Louis Pasteur, Strasbourg Cedex, France
Laboratoire d'Hématologie, Hôpital Hautepierre, Strasbourg, France
Additional corresponding author
Search for more papers by this authorAbstract
Shed membrane microparticles (MPs) are microvesicles generated from the plasma membrane when cells are submitted to stress conditions. Although MPs reflect the cell state (at least in vitro), little is known on their protein composition. We describe the first set of experiments aiming to characterize the MP proteome. Two ways of triggering MP formation from a T-lymphocytic cell line were analyzed using a 1-D gel approach coupled with LC-MS/MS and the results were compared with those obtained from a classic membrane preparation. In total, 390 proteins were identified in MPs, among which 34% were localized to the plasma membrane. The MPs revealed a broad representation of plasma membrane proteins including 17 hematopoietic clusters of differentiation. This approach was successfully applied to one human chronic B-cell lymphoid malignancy. In all, 413 proteins were identified, including 117 membrane proteins, many of them being pathology associated. The sequence coverage in identified proteins was improved combining both nano-LC-MS/MS and MALDI-MS data. The suppression effect, observed on very complex peptide mixtures, was remediated by chromatographic fractionation. MPs may represent a new tool for studying plasma membrane proteins, displaying the advantages of reproducibility, minimal organelle contamination, and being potentially applicable to most cell types.
Supporting Information
Supporting information for this article is available on the WWW under http://www.wiley-vch.de/contents/jc_2120/2006/pro0133_s.pdf or from the author.
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