Assembly and Ligand Binding Properties of the Water-soluble Extracellular Domains of the Glutamate Receptor 1 Subunit

High resolution structural studies of models of glutamate receptors (GluRs) have been limited to monomeric models of the ligand-binding site. To obtain oligomeric models of glutamate receptors that can reveal more complete structural information, we examined the assembly and ligand binding propertie...

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Bibliographic Details
Published in:The Journal of biological chemistry 2001-02, Vol.276 (5), p.3031-3036
Main Authors: Wells, Gregg B., Lin, Lin, Jeanclos, Elisabeth M., Anand, Rene
Format: Article
Language:English
Subjects:
alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid - metabolism
Animals
Ligands
Membrane Proteins - chemistry
Membrane Proteins - metabolism
Mutagenesis, Site-Directed
Oocytes - metabolism
Protein Conformation
Protein Structure, Tertiary
Receptors, AMPA - chemistry
Receptors, AMPA - genetics
Receptors, AMPA - metabolism
Solubility
Transfection
Tritium
Water - chemistry
Xenopus laevis
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Summary:High resolution structural studies of models of glutamate receptors (GluRs) have been limited to monomeric models of the ligand-binding site. To obtain oligomeric models of glutamate receptors that can reveal more complete structural information, we examined the assembly and ligand binding properties of two truncated versions of the GluR1 subunit. The first version, GluR1-WS, consisted of only the N-terminal extracellular segment (Ala1–Glu520) bridged by a synthetic linker to the second extracellular domain (Asn615–Gly790). The second version, GluR1-M1, consisted of the first N-terminal extracellular domain (Ala1–Glu520) bridged by a synthetic linker to a second segment containing the second extracellular domain, the third transmembrane domain, and the intracellular C-terminal domain (Asn615–Leu889). When expressed inXenopus oocytes, GluR-WS was secreted and water-soluble; GluR1-M1 was displayed on the surface of oocytes. GluR1-WS exhibited a velocity sedimentation profile that was consistent with assembly of homooligomers and bound the glutamate receptor agonist α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid with high affinity. These findings show that the extracellular domains of GluR1 that are sufficient for ligand binding apparently are sufficient for subunit assembly and might be a suitable target for structural studies of a water-soluble GluR1 oligomer.
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.M006668200