Interactions of the C Terminus of Metabotropic Glutamate Receptor Type 1α with Rat Brain Proteins

Interactions of the C Terminus of Metabotropic Glutamate Receptor Type 1α with Rat Brain Proteins Abstract : Metabotropic glutamate receptors (mGluRs) are coupled to G protein second messenger pathways and modulate glutamate neurotransmission in the brain, where they are targeted to specific synaptic locations. As part of a strategy for defining the mechanisms for the specific targeting of mGluR1 α, rat brain proteins which interact with the intracellular carboxy terminus of mGluR1 α have been characterized, using affinity chromatography on a glutathione S‐transferase fusion protein that contains the last 86 amino acids of mGluR1 α. Three of the proteins specifically eluted from the affinity column yielded protein sequences, two of which were identified as glyceraldehyde‐3‐phosphate dehydrogenase and β‐tubulin ; the other was an unknown protein. The identity of tubulin was confirmed by western immunoblotting. Using a solid‐phase binding assay, the mGluR1 α‐tubulin interaction was shown to be direct, specific, and saturable with a KD of 2.3 ± 0.4 μM. In addition, mGluR1 α, but not mGluR2/3 or mGluR4, could be coimmunoprecipitated from solubilized brain extracts with tubulin using anti‐β‐tubulin antibodies. However, mGluR1 α could not be coimmunoprecipitated with the tubulin binding protein gephyrin, nor could it be coimmunoprecipitated with PSD95. Collectively these data demonstrate that the last 86 amino acids of the carboxyl‐terminal tail of mGluR1 α are sufficient to determine its interaction with tubulin and that there is an association of this receptor with tubulin in rat brain. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Neurochemistry Wiley

Interactions of the C Terminus of Metabotropic Glutamate Receptor Type 1α with Rat Brain Proteins

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Publisher
Wiley Subscription Services, Inc., A Wiley Company
Copyright
© International Society for Neurochemistry
ISSN
0022-3042
eISSN
1471-4159
D.O.I.
10.1046/j.1471-4159.1999.0720346.x
Publisher site
See Article on Publisher Site

Abstract

Abstract : Metabotropic glutamate receptors (mGluRs) are coupled to G protein second messenger pathways and modulate glutamate neurotransmission in the brain, where they are targeted to specific synaptic locations. As part of a strategy for defining the mechanisms for the specific targeting of mGluR1 α, rat brain proteins which interact with the intracellular carboxy terminus of mGluR1 α have been characterized, using affinity chromatography on a glutathione S‐transferase fusion protein that contains the last 86 amino acids of mGluR1 α. Three of the proteins specifically eluted from the affinity column yielded protein sequences, two of which were identified as glyceraldehyde‐3‐phosphate dehydrogenase and β‐tubulin ; the other was an unknown protein. The identity of tubulin was confirmed by western immunoblotting. Using a solid‐phase binding assay, the mGluR1 α‐tubulin interaction was shown to be direct, specific, and saturable with a KD of 2.3 ± 0.4 μM. In addition, mGluR1 α, but not mGluR2/3 or mGluR4, could be coimmunoprecipitated from solubilized brain extracts with tubulin using anti‐β‐tubulin antibodies. However, mGluR1 α could not be coimmunoprecipitated with the tubulin binding protein gephyrin, nor could it be coimmunoprecipitated with PSD95. Collectively these data demonstrate that the last 86 amino acids of the carboxyl‐terminal tail of mGluR1 α are sufficient to determine its interaction with tubulin and that there is an association of this receptor with tubulin in rat brain.

Journal

Journal of NeurochemistryWiley

Published: Jan 1, 1999

Keywords: ; ; ;

References

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