Molecular Cloning, Expression, and Pharmacological Characterization of humEAA1, a Human Kainate Receptor Subunit
| Autor: | Darryle D. Schoepp, Stephen L. Nutt, Wosnick Michael A, B. Korczak, Vikarna Rampersad, Rajender Kamboj, Rebecca A. True, Dennis M. Zimmerman, Shekter Lee |
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| Rok vydání: | 2008 |
| Předmět: |
Kainic acid
DNA Complementary Macromolecular Substances Protein subunit Molecular Sequence Data Gene Expression Kainate receptor Biology Binding Competitive Polymerase Chain Reaction Biochemistry Mice Cellular and Molecular Neuroscience chemistry.chemical_compound Receptors Kainic Acid Complementary DNA Animals Humans Amino Acid Sequence Cloning Molecular Receptor DNA Primers Kainic Acid Base Sequence Sequence Homology Amino Acid Chinese hamster ovary cell Glutamate receptor Molecular biology Rats Kinetics chemistry NMDA receptor Chickens |
| Zdroj: | Journal of Neurochemistry. 62:1-9 |
| ISSN: | 1471-4159 0022-3042 |
| Popis: | Kainate is a potent neuroexcitatory agent; its neurotoxicity is thought to be mediated by an ionotropic receptor with a nanomolar affinity for kainate. In this report, we describe the cloning of a cDNA encoding a human glutamate ionotropic receptor subunit protein from a human hippocampal library. This cDNA, termed humEAA1, is most closely related to rat and human cDNAs for kainate receptor proteins and, when expressed in COS or Chinese hamster ovary cells, is associated with high-affinity kainate receptor binding. We have successfully established cell lines stably expressing humEAA1. This is the first report of establishment of stable cell lines expressing a glutamate receptor subunit. The relative potency of compounds for displacing [3H] kainate binding of humEAA1 receptors expressed in these stable cell lines was kainate > quisqualate > domoate > L-glutamate > (RS)-α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid > dihydro-kainate > 6, 7-dinitroquinoxaline-2, 3-dione > 6-cyano-7-nitroquinoxaline-2, 3-dione. Homooligomeric expression of humEAA1 does not appear to elicit ligand-gated ion channel activity. Nevertheless, the molecular structure and pharmacological characterization of high-affinity kainate binding of the humEAA1 expressed in the stable cell line (ppEAA1–16) suggest that the humEAA1 is a subunit protein of a human kainate receptor complex. |
| Databáze: | OpenAIRE |
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