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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.
- Source :
-
Molecular and cellular neurosciences [Mol Cell Neurosci] 2001 Sep; Vol. 18 (3), pp. 296-306. - Publication Year :
- 2001
-
Abstract
- 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.<br /> (Copyright 2001 Academic Press.)
- Subjects :
- Animals
CHO Cells drug effects
CHO Cells metabolism
Carrier Proteins biosynthesis
Cricetinae
Dimerization
Dose-Response Relationship, Drug
Excitatory Amino Acid Antagonists pharmacology
Green Fluorescent Proteins
Homer Scaffolding Proteins
Humans
Immunoblotting
Neuropeptides biosynthesis
Precipitin Tests
Pyridines pharmacology
Receptor, Metabotropic Glutamate 5
Receptors, Metabotropic Glutamate antagonists & inhibitors
Receptors, Metabotropic Glutamate biosynthesis
Recombinant Fusion Proteins antagonists & inhibitors
Recombinant Fusion Proteins biosynthesis
Signal Transduction drug effects
Signal Transduction physiology
Transfection methods
Carrier Proteins physiology
Luminescent Proteins genetics
Neuropeptides physiology
Receptors, Metabotropic Glutamate genetics
Receptors, Metabotropic Glutamate metabolism
Recombinant Fusion Proteins genetics
Recombinant Fusion Proteins metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1044-7431
- Volume :
- 18
- Issue :
- 3
- Database :
- MEDLINE
- Journal :
- Molecular and cellular neurosciences
- Publication Type :
- Academic Journal
- Accession number :
- 11591130
- Full Text :
- https://doi.org/10.1006/mcne.2001.1031