Domain organization and function in GluK2 subtype kainate receptors.
Glutamate receptor ion channels (iGluRs) are excitatory neurotransmitter receptors with a unique molecular architecture in which the extracellular domains assemble as a dimer of dimers. The structure of individual dimer assemblies has been established previously for both the isolated ligand-binding domain (LBD) and more recently for the larger amino terminal domain (ATD). How these dimers pack to form tetrameric assemblies in intact iGluRs has remained controversial. Using recently solved crystal structures for the GluK2 kainate receptor ATD as a guide, we performed cysteine mutant cross-linking experiments in full-length tetrameric GluK2 to establish how the ATD packs in a dimer of dimers assembly. A similar approach, using a full-length AMPA receptor GluA2 crystal structure as a guide, was used to design cysteine mutant cross-links for the GluK2 LBD dimer of dimers assembly. The formation of cross-linked tetramers in full-length GluK2 by combinations of ATD and LBD mutants which individually produce only cross-linked dimers suggests that subunits in the ATD and LBD layers swap dimer partners. Functional studies reveal that cross-linking either the ATD or the LBD inhibits activation of GluK2 and that, in the LBD, cross-links within and between dimers have different effects. These results establish that kainate and AMPA receptors have a conserved extracellular architecture and provide insight into the role of individual dimer assemblies in activation of ion channel gating.
Das
Utpal
U
Laboratory of Cellular and Molecular Neurophysiology, Department of Health and Human Services, Porter Neuroscience Research Center, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA.
Kumar
Janesh
J
Mayer
Mark L
ML
Plested
Andrew J R
AJ
D000818:Animals; D002460:Cell Line; D018360:Crystallography, X-Ray; D006801:Humans; D015640:Ion Channel Gating; D008024:Ligands; D008958:Models, Molecular; D009154:Mutation; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D051381:Rats; D018092:Kainic Acid Receptors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000097806:GluK2 Kainate Receptor
D008024:Ligands; D018092:Kainic Acid Receptors; D000097806:GluK2 Kainate Receptor
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