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Updated: Aug 14, 2026
Quantitative Mass Spectrometric Profiling of Cancer-cell Proteomes Derived From Liquid and Solid Tumors
Published on: February 27, 2015
Proteomic analysis of malignant lymphocyte membrane microparticles using double ionization coverage optimization
Laurent Miguet1, Karine Pacaud, Claire Felden
1Laboratoire de Spectrométrie de Masse Bio-Organique, Université Louis Pasteur, CNRS UMR 7512, 25 rue Becquerel, 67087 Strasbourg Cedex 2, France.
Abstract:
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.
Insights
Researchers characterized the protein composition of shed membrane microparticles (MPs), which originate from cell membranes under stress. This proteomic analysis reveals MPs as a valuable tool for studying plasma membrane proteins.
Area of Science:
- Proteomics
- Cell Biology
- Biochemistry
Background:
- Shed membrane microparticles (MPs) are vesicles released from the plasma membrane during cellular stress.
- While MPs reflect cellular states, their protein composition remains largely uncharacterized.
- Understanding MP protein content is crucial for their application in biological research.
Purpose of the Study:
- To characterize the proteome of shed membrane microparticles (MPs).
- To establish a method for analyzing MP protein composition.
- To evaluate the potential of MPs as tools for studying plasma membrane proteins.
Main Methods:
- MP formation was induced in a T-lymphocytic cell line under stress conditions.
- Proteomic analysis was performed using 1-D gel electrophoresis coupled with LC-MS/MS.
- MP protein identification was compared to classic membrane preparations and validated with MALDI-MS.
Main Results:
- A total of 390 proteins were identified in MPs from the T-lymphocytic cell line, with 34% localized to the plasma membrane.
- MPs showed a diverse representation of plasma membrane proteins, including hematopoietic clusters of differentiation.
- The method was successfully applied to a human chronic B-cell lymphoid malignancy, identifying 413 proteins, including 117 membrane proteins.
Conclusions:
- Membrane microparticles (MPs) offer a reproducible method for studying plasma membrane proteins with minimal organelle contamination.
- The proteomic characterization of MPs provides insights into cellular stress responses and potential disease biomarkers.
- MPs are a promising tool applicable to most cell types for proteomic analysis.
