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Figure 1. Sequence alignments for S1 and S2 regions in NR2A, NR2B, NR2C and NR2D NMDAR subunits. Residues shown in blue are common to all four NMDAR subunits. Residues shown in red are common to three subunits, while those shown in bold black are either different in each of the four subunits or are common to only two subunits. The two locations in the S2 region where residues are conserved in NR2B, NR2C and NR2D NMDAR subunits but are different in the NR2A NMDAR subunit and result in a change in charge at such locations are shown in bold red. These are amino acids Lys719 (NR2A), Met713 (NR2B), Met717 (NR2C) and Met740 (NR2D) and Tyr735 (NR2A), Lys729 (NR2B), Lys733 (NR2C) and Lys756 (NR2D).

Figure 2. Glycine concentration-response data for wild-type NR2A, NR2D and NR2A(2D-S1S2) NMDARs when homoquinolinate is used as the NR2 binding site agonist. The use of homoquinolinate (rather than glutamate) as the NR2 agonist does not alter the potency of glycine. EC50 values for glycine acting at each of these NMDAR combinations are given in the text. The mean Hill slopes and mean maximal currents recorded were for NR2A(WT): 1.17 ± 0.07, 3.5 ± 0.4 μA; NR2D(WT): 1.19 ± 0.06, 0.4 ± 0.06 μA and NR2A(2D-S1S2): 1.37 ± 0.06, 1.4 ± 0.7 μA.

Figure 3. Concentration-response curves for chimeric NMDARs where membrane associated domains and their intracellular linkers from NR2A have been replaced with those found in NR2D subunits. A, mean glutamate concentration-response curves for NR2A(2D-M1M2M3) and NR2A(2D-S1M1M2M3S2) constructs. Inclusion of the M1, M2 and M3 domains from the NR2D subunit in NR2A subunits does not alter glutamate potency compared to NR2A(WT)-containing NMDARs. When S1 and S2 regions are also included a shift to a NR2D-like potency is observed. For comparison, the dashed lines show the comparable curves for NR2A(WT)- and NR2D(WT)- containing NMDARs (from Erreger et al. 2007). For glutamate concentration-response curves, mean Hill slopes and mean maximal currents recorded were for NR2A(2DM1M2M3): 1.18 ± 0.04, 2.0 ± 0.1 μA and NR2A(2D-S1M1M2M3S2): 1.23 ± 0.05, 1.1 ± 0.2 μA. B, mean glycine concentration-response curves for NR2A(2D-M1M2M3) and NR2A(2D-S1M1M2M3S2) constructs. Inclusion of the M1, M2 and M3 domains from the NR2D subunit in NR2A subunits does not alter glycine potency compared to NR2A(WT)-containing NMDARs. When S1 and S2 regions are also included a shift to a more NR2D-like potency is observed. For comparison, the dashed lines show the comparable curves for NR2A(WT)- and NR2D(WT)-containing NMDARs. For glycine concentration-response curves, mean Hill slopes and mean maximal currents recorded were for NR2A(2D-M1M2M3): 1.30 ± 0.06, 1.4 ± 0.2 μA and NR2A(2D-S1M1M2M3S2): 1.14 ± 0.04, 0.5 ± 0.05 μA.

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