Characterization of a Metabotropic Glutamate Receptor Type 5-Green Fluorescent Protein Chimera (mGluR5–GFP): Pharmacology, Surface Expression, and Differential Effects of Homer-1a and Homer-1c

Autor: Michael A. Tones, Victoria Coutinho, Irene Kavanagh, Jeremy M. Henley, Hiroyuki Sugiyama
Rok vydání: 2001
Předmět:
Pyridines
Receptor
Metabotropic Glutamate 5
Recombinant Fusion Proteins
animal diseases
Green Fluorescent Proteins
Immunoblotting
CHO Cells
Biology
Receptors
Metabotropic Glutamate
Transfection
Cellular and Molecular Neuroscience
Homer Scaffolding Proteins
Cricetinae
mental disorders
Animals
Humans
Molecular Biology
Dose-Response Relationship
Drug
Metabotropic glutamate receptor 5
Metabotropic glutamate receptor 4
Neuropeptides
fungi
Metabotropic glutamate receptor 7
Metabotropic glutamate receptor 6
Cell Biology
Precipitin Tests
Cell biology
Luminescent Proteins
nervous system
Metabotropic glutamate receptor
Metabotropic glutamate receptor 1
Metabotropic glutamate receptor 3
Metabotropic glutamate receptor 2
Carrier Proteins
Dimerization
Excitatory Amino Acid Antagonists
Signal Transduction
Zdroj: Molecular and Cellular Neuroscience. 18:296-306
ISSN: 1044-7431
Popis: Metabotropic glutamate receptor 5 (mGluR5) can modulate synaptic transmission by increasing intracellular Ca2+ and it plays a role in several forms of synaptic plasticity. We have constructed a fusion of human mGluR5 and green fluorescent protein (mGluR5-GFP). Expression of mGluR5-GFP in clonal cell lines yielded a functional fluorescent receptor with pharmacological profiles similar to wild-type mGluR5. mGluR5-GFP coimmunoprecipitated with Homer-1c, indicating that addition of GFP to the C-terminal did not prevent Homer binding. Coexpression of wild-type mGluR5 or mGluR5-GFP with Homer 1c, but not Homer-1a, resulted in reduced receptor surface localization and the formation of intracellular clusters. Neither Homer-1a nor Homer-1c had any effect on mGluR1 or mGluR1-GFP distribution. mGluR5-GFP expressed alone or in combination with Homer-1a formed dimers in HEK cells. Coexpression with Homer-1c, however, prevented mGluR5-GFP dimerization. Neither Homer altered the agonist profiles of mGluR5 or mGluR5-GFP. These data indicate that the functional expression of mGluR5 is regulated by Homer-1c and demonstrate that mGluR5-GFP provides a useful tool to study the molecular pharmacology and cell biology of mGluRs in real-time.
Databáze: OpenAIRE
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