Single-channel properties of N-methyl-D-aspartate receptors containing chimaeric GluN2A/GluN2D subunits.
Subtypes of NMDARs (N-methyl-D-aspartate receptors) display differences in their pharmacological and biophysical properties. The differences are, to a large extent, determined by the identities of the GluN2 (glutamate-binding) NMDAR subunits that are co-expressed with GluN1 (glycine-binding) subunits, which form the final tetrameric NMDAR assembly. Of the four GluN2 subunits that exist (termed A-D), NMDARs composed of GluN1/GluN2A and GluN1/GluN2D subunits display the greatest differences in their sensitivities to a variety of agonists, antagonists and channel blockers as well as showing marked differences in their single-channel conductances and deactivation kinetics. Here, we describe a series of experiments where we have generated and studied two chimaeric GluN2A/GluN2D subunits. The first chimaera, referred to as GluN2A(2D-M1M2M3), replaces the membrane-associated regions M1, M2 and M3 of the GluN2A subunit with the corresponding regions found in the GluN2D subunit. The second chimaera, GluN2A(2D-S1M1M2M3S2), replaces the same three membrane-associated regions of the GluN2A subunit plus the LBD (ligand-binding domain) with the corresponding regions of the GluN2D subunit. Our results show that the identity of the GluN2 LBD not only controls glutamate potency, but also influences the potency of the NMDAR co-agonist glycine, whereas the single-channel conductance and the duration of single activations of ion channels can be predicted by the identities of the M1-M3 regions and the LBD.
O'Leary
Timothy
T
Centre for Integrative Physiology, University of Edinburgh, Hugh Robson Building, George Square, Edinburgh EH8 9XD, UK.
Wyllie
David J A
DJ
D000818:Animals; D004594:Electrophysiology; D018698:Glutamic Acid / Q000378:metabolism; D005998:Glycine / Q000378:metabolism; D008274:Magnesium / Q000378:metabolism; D008958:Models, Molecular; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016194:Receptors, N-Methyl-D-Aspartate / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D021122:Protein Subunits; D016194:Receptors, N-Methyl-D-Aspartate; D011993:Recombinant Fusion Proteins; D018698:Glutamic Acid; D008274:Magnesium; D005998:Glycine