Abstract
The pathological importance of tumor necrosis factor (TNF)-α in rheumatoid arthritis (RA) is now widely accepted. Ex vivo data from synovial cell cultures suggest that direct cell contact between activated T-cells and macrophages may be an important driver of macrophage TNF-α production in the RA joint. However, the ligand/receptor pairs driving this cell contact signal remain obscure. One reason for this is that plasma membrane (PM) proteins are resistant to systematic analysis using traditional proteomic approaches. In this chapter we present a method for the enrichment and resolution of PM proteins from murine T-cell hybridomas as a prelude to identification by tandem mass spectrometry. We used cell surface biotinylation, differential centrifugation and subsequent streptavidin affinity capture, followed by solution phase iso-electric focussing and tandem mass spectrometry to identify 75 PM proteins and make semiquantitative comparisons of resting and activated cells. The method is applicable to a wide variety of cell types.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Li, J. M., Isler, P., Dayer, J. M., and Burger, D. (1995) Contact-dependent stimulation of monocytic cells and neutrophils by stimulated human T-cell clones. Immunology 84, 571–576.
McInnes, I. B., Leung, B. P., Sturrock, R. D., Field, M., and Liew, F. Y. (1997) Interleukin-15 mediates T cell-dependent regulation of tumor necrosis factor-alpha production in rheumatoid arthritis. Nat. Med. 3, 189–195.
Brennan, F. M., Hayes, A. L., Ciesielski, C. J., Green, P., Foxwell, B. M., and Feldmann, M. (2002) Evidence that rheumatoid arthritis synovial T cells are similar to cytokine-activated T cells: involvement of phosphatidylinositol 3-kinase and nuclear factor kappaB pathways in tumor necrosis factor alpha production in rheumatoid arthritis. Arthritis Rheum. 46, 31–41.
Vey, E., Dayer, J. M., and Burger, D. (1997) Direct contact with stimulated T cells induces the expression of IL-1beta and IL-1 receptor antagonist in human monocytes. Involvement of serine/threonine phosphatases in differential regulation. Cytokine 9, 480–487.
Lacraz, S., Isler, P., Vey, E., Welgus, H. G., and Dayer, J. M. (1994) Direct contact between T lymphocytes and monocytes is a major pathway for induction of metalloproteinase expression. J. Biol. Chem. 269, 22,027–22,033.
Ribbens, C., Dayer, J. M., and Chizzolini, C. (2000) CD40-CD40 ligand (CD154) engagement is required but may not be sufficient for human T helper 1 cell induction of interleukin-2-or interleukin-15-driven, contact-dependent, interleukin-1beta production by monocytes. Immunology 99, 279–286.
Isler, P., Vey, E., Zhang, J. H., and Dayer, J. M. (1993) Cell surface glycoproteins expressed on activated human T cells induce production of interleukin-1 beta by monocytic cells: a possible role of CD69. Eur. Cytokine Netw. 4, 15–23.
Santoni, V., Molloy, M., and Rabilloud, T. (2000) Membrane proteins and proteomics: un amour impossible? Electrophoresis 21, 1054–1070.
Gygi, S. P., Corthals, G. L., Zhang, Y., Rochon, Y., and Aebersold, R. (2000). Evaluation of two-dimensional gel electrophoresis-based proteome analysis technology. Proc. Natl. Acad. Sci. USA 97, 9390–9395.
Peirce, M. J., Wait, R., Begum, S., Saklatvala, J., and Cope, A. P. (2004). Expression profiling of lymphocyte plasma membrane proteins. Mol. Cell Proteomics 3, 56–65.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2007 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Peirce, M.J., Saklatvala, J., Cope, A.P., Wait, R. (2007). Mapping Lymphocyte Plasma Membrane Proteins. In: Cope, A.P. (eds) Arthritis Research. Methods in Molecular Medicine, vol 136. Humana Press. https://doi.org/10.1007/978-1-59745-402-5_25
Download citation
DOI: https://doi.org/10.1007/978-1-59745-402-5_25
Publisher Name: Humana Press
Print ISBN: 978-1-58829-918-5
Online ISBN: 978-1-59745-402-5
eBook Packages: Springer Protocols