Proteomic analysis of native metabotropic glutamate receptor 5 protein complexes reveals novel molecular constituents

Carol D. Farr, Philip R. Gafken, Angela D. Norbeck, Catalin E. Doneanu, Martha D. Stapels, Douglas F. Barofsky, Manabu Minami, Julie Saugstad

Research output: Contribution to journalArticle

60 Scopus citations

Abstract

We used a proteomic approach to identify novel proteins that may regulate metabotropic glutamate receptor 5 (mGluRS) 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.

Original languageEnglish (US)
Pages (from-to)438-450
Number of pages13
JournalJournal of Neurochemistry
Volume91
Issue number2
DOIs
Publication statusPublished - Oct 2004
Externally publishedYes

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Keywords

  • Mass spectrometry
  • Metabotropic glutamate receptor
  • Protein interaction
  • Proteomics

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

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