- Open Access
A proteomic approach based on peptide affinity chromatography, 2-dimensional electrophoresis and mass spectrometry to identify multiprotein complexes interacting with membrane-bound receptors
Biological Procedures Online volume 4, pages 94–104 (2002)
There is accumulating evidence that membrane-bound receptors interact with many intracellular proteins. Multiprotein complexes associated with ionotropic receptors have been extensively characterized, but the identification of proteins interacting with G protein-coupled receptors (GPCRs) has so far only been achieved in a piecemeal fashion, focusing on one or two protein species. We describe a method based on peptide affinity chromatography, two-dimensional electrophoresis, mass spectrometry and immunoblotting to identify the components of multiprotein complexes interacting directly or indirectly with intracellular domains of GPCRs or, more generally, any other membrane-bound receptor. Using this global approach, we have characterized multiprotein complexes that bind to the carboxy-terminal tail of the 5-hydroxytryptamine type 2C receptor and are important for its subcellular localization in CNS cells (Bécamel et al., EMBO J., 21(10): 2332, 2002).
Husi H, Ward MA, Choudhary JS, Blackstock WP, Grant SG. Proteomic analysis of NMDA receptor-adhesion protein signaling complexes. Nat Neurosci 2000;3:661–669.
Kim M, Jiang LH, Wilson HL, North RA, Surprenant A. Proteomic and functional evidence for a P2X(7) receptor signalling complex. Embo J 2001;20:6347–6358.
Brzostowski JA, Kimmel AR. Signaling at zero G: G-protein-independent functions for 7-TM receptors. Trends Biochem Sci 2001;26:291–297.
Milligan G, White J. Protein-protein interactions at G-protein-coupled receptors. Trends Pharmacol Sci 2001;22:513–518.
Brady AE, Limbird LE. G protein-coupled receptor interacting proteins: emerging roles in localization and signal transduction. Cell Signal 2002;14:297–309.
Bockaert J, Claeysen S, Bécamel C, Pinloche S, Dumuis A. G protein-coupled receptors: dominant players in cell-cell communication. Int Rev Cytol 2002;212:63–132.
Sheng M, Sala C. PDZ domains and the organization of supramolecular complexes. Annu Rev Neurosci 2001;24:1–29.
Hall RA, Premont RT, Chow CW, Blitzer JT, Pitcher JA, Claing A, Stoffel RH, Barak LS, Shenolikar S, Weinman EJ, Grinstein S, Lefkowitz RJ. The beta2- adrenergic receptor interacts with the Na+/H+-exchanger regulatory factor to control Na+/H+ exchange. Nature 1998;392:626–630.
Ullmer C, Schmuck K, Figge A, Lubbert H. Cloning and characterization of MUPP1, a novel PDZ domain protein. FEBS Lett 1998;424:63–68.
Zitzer H, Honck HH, Bachner D, Richter D, Kreienkamp HJ. Somatostatin receptor interacting protein defines a novel family of multidomain proteins present in human and rodent brain. J Biol Chem 1999b;274:32997–33001.
Raymond JR, Mukhin YV, Gelasco A, Turner J, Collinsworth G, Gettys TW, Grewal JS, Garnovskaya MN. Multiplicity of mechanisms of serotonin receptor signal transduction. Pharmacol Ther 2001;92:179–212.
Bécamel C, Alonso G, Galeotti N, Demey E, Jouin P, Ullmer C, Dumuis A, Bockaert J, Marin P. Synaptic multiprotein complexes associated with 5-HT(2C) receptors: a proteomic approach. Embo J 2002;21:2332–2342.
Misawa H, Kawasaki Y, Mellor J, Sweeney N, Jo K, Nicoll RA, Bredt DS. Contrasting localizations of MALS/LIN-7 PDZ proteins in brain and molecular compensation in knockout mice. J Biol Chem 2001;276:9264–9272.
Bécamel C, Figge A, Poliak S, Dumuis A, Peles E, Bockaert J, Lubbert H, Ullmer C. Interaction of serotonin 5-hydroxytryptamine type 2C receptors with PDZ10 of the multi-PDZ domain protein MUPP1. J Biol Chem 2001;276:12974–12982.
Laoudj-Chenivesse D, Marin P, Bennes R, Tronel-Peyroz E, Leterrier F. High performance two-dimensional gel electrophoresis using a wetting agent Tergitol(R) NP7. Proteomics 2002;2:481–485.
Shevchenko A, Wilm M, Vorm O, Mann M. Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal Chem 1996;68:850–858.
Lee K, Bae D, Lim D. Evaluation of parameters in peptide mass fingerprinting for protein identification by MALDITOF mass spectrometry. Mol Cells 2002;13:175–184.
Karas M, Hillenkamp F. Laser desorption ionization of proteins with molecular masses exceeding 10,000 daltons. Anal Chem 1988;60:2299–2301.
Garin J, Diez R, Kieffer S, Dermine JF, Duclos S, Gagnon E, Sadoul R, Rondeau C, Desjardins M. The phagosome proteome: insight into phagosome functions. J Cell Biol 2001;152:165–180.
Mann M, Wilm M. Error-tolerant identification of peptides in sequence databases by peptide sequence tags. Anal Chem 1994;66:4390–4399.
Mann M, Hendrickson RC, Pandey A. Analysis of Proteins and Proteomes by Mass Spectrometry. Annu Rev Biochem 2001;70:437–473.
Ficarro SB, McCleland ML, Stukenberg PT, Burke DJ, Ross MM, Shabanowitz J, Hunt DF, White FM. Phosphoproteome analysis by mass spectrometry and its application to Saccharomyces cerevisiae. Nat Biotechnol 2002;20:301–305.
Jo K, Derin, R, Li, M, Bredt, DS. Characterization of MALS/Velis-1, -2, and -3: a family of mammalian LIN- 7 homologs enriched at brain synapses in association with the postsynaptic density-95/NMDA receptor postsynaptic complex. J Neurosci 1999;19:4189–4199.
Butz S, Okamoto, M, Südhof, TC. A tripartite protein complex with the potential to couple synaptic vesicle exocytosis to cell adhesion in brain. Cell 1998;94:773–782.
Borg JP, Straight, SW, Kaech, SM, de Taddeo-Borg, M, Kroon, DE, Karnak, D, Turner, RS, Kim, SK, Margolis, B. Identification of an evolutionarily conserved heterotrimeric protein complex involved in protein targeting. J Biol Chem 1998;273:31633–31636.
Published: December 9, 2002
About this article
Cite this article
Bécamel, C., Galéotti, N., Poncet, J. et al. A proteomic approach based on peptide affinity chromatography, 2-dimensional electrophoresis and mass spectrometry to identify multiprotein complexes interacting with membrane-bound receptors. Biol Proced Online 4, 94–104 (2002). https://doi.org/10.1251/bpo39
- spectrum analysis, mass