Proteomic analysis of native metabotropic glutamate receptor 5 protein complexes reveals novel molecular constituents
Open Access
- 14 September 2004
- journal article
- Published by Wiley in Journal of Neurochemistry
- Vol. 91 (2), 438-450
- https://doi.org/10.1111/j.1471-4159.2004.02735.x
Abstract
We used a proteomic approach to identify novel proteins that may regulate metabotropic glutamate receptor 5 (mGluR5) responses by direct or indirect protein interactions. This approach does not rely on the heterologous expression of proteins and offers the advantage of identifying protein interactions in a native environment. The mGluR5 protein was immunoprecipitated from rat brain lysates; co‐immunoprecipitating proteins were analyzed by mass spectrometry and identified peptides were matched to protein databases to determine the correlating parent proteins. This proteomic approach revealed the interaction of mGluR5 with known regulatory proteins, as well as novel proteins that reflect previously unidentified molecular constituents of the mGluR5‐signaling complex. Immunoblot analysis confirmed the interaction of high confidence proteins, such as phosphofurin acidic cluster sorting protein 1, microtubule‐associated protein 2a and dynamin 1, as mGluR5‐interacting proteins. These studies show that a proteomic approach can be used to identify candidate interacting proteins. This approach may be particularly useful for neurobiology applications where distinct protein interactions within a signaling complex can dramatically alter the outcome of the response to neurotransmitter release, or the disruption of normal protein interactions can lead to severe neurological and psychiatric disorders.Keywords
This publication has 75 references indexed in Scilit:
- The distinct role of mGlu1 receptors in post-ischemic neuronal deathTrends in Pharmacological Sciences, 2003
- The Metabotropic Glutamate Receptor mGluR5 Is Endocytosed by a Clathrin-independent PathwayJournal of Biological Chemistry, 2003
- Structural, signalling and regulatory properties of the group I metabotropic glutamate receptors: prototypic family C G-protein-coupled receptorsBiochemical Journal, 2001
- Agonist‐induced internalization of the metabotropic glutamate receptor 1a is arrestin‐ and dynamin‐dependentJournal of Neurochemistry, 2001
- Endogenous sulphur‐containing amino acids: potent agonists at presynaptic metabotropic glutamate autoreceptors in the rat central nervous systemBritish Journal of Pharmacology, 2001
- The Millennium of the Dendrite?Neuron, 2000
- Homer 1b Regulates the Trafficking of Group I Metabotropic Glutamate ReceptorsPublished by Elsevier ,1999
- Regulation of Endosome Sorting by a Specific PP2A IsoformThe Journal of cell biology, 1998
- Identification of the Binding Site for Acidic Phospholipids on the PH Domain of Dynamin: Implications for Stimulation of GTPase ActivityJournal of Molecular Biology, 1996
- An approach to correlate tandem mass spectral data of peptides with amino acid sequences in a protein databaseJournal of the American Society for Mass Spectrometry, 1994