Proteomic analysis of native metabotropic glutamate receptor 5 protein complexes reveals novel molecular constituents
Article first published online: 14 SEP 2004
Journal of Neurochemistry
Volume 91, Issue 2, pages 438–450, October 2004
How to Cite
Farr, C. D., Gafken, P. R., Norbeck, A. D., Doneanu, C. E., Stapels, M. D., Barofsky, D. F., Minami, M. and Saugstad, J. A. (2004), Proteomic analysis of native metabotropic glutamate receptor 5 protein complexes reveals novel molecular constituents. Journal of Neurochemistry, 91: 438–450. doi: 10.1111/j.1471-4159.2004.02735.x
- Issue published online: 14 SEP 2004
- Article first published online: 14 SEP 2004
- Received April 5, 2004; revised manuscript received June 25, 2004; accepted June 29, 2004.
- mass spectrometry;
- metabotropic glutamate receptor;
- protein interaction;
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.