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Proteomic analysis of native metabotropic glutamate receptor 5 protein complexes reveals novel molecular constituents
Author(s) -
Farr Carol D.,
Gafken Philip R.,
Norbeck Angela D.,
Doneanu Catalin E.,
Stapels Martha D.,
Barofsky Douglas F.,
Minami Manabu,
Saugstad Julie A.
Publication year - 2004
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.2004.02735.x
Subject(s) - metabotropic glutamate receptor 5 , protein–protein interaction , metabotropic glutamate receptor , biology , metabotropic glutamate receptor 6 , interactome , immunoprecipitation , metabotropic glutamate receptor 1 , microbiology and biotechnology , proteomics , glutamate receptor , chemistry , biochemistry , receptor , gene
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.