Table 1.
Effects of ABD dimer interface mutations on Zn2+ and H+ inhibitions of NR1/NR2A receptors
| Mutant | Zn2+ inhibition |
H+ inhibition |
|||||
|---|---|---|---|---|---|---|---|
| IC50 (nM) | Max. Inhib. (%) | nH | # cells | pH IC50 | nH | # cells | |
| wild-type | 10.5 ± 0.8 | 81 ± 1 | 0.91 | (5) | 6.95 ± 0.05 | 1.69 | (14) |
| NR1 mutant | |||||||
| I519A | 8.9 ± 2.1 | 84 ± 1 ** | 0.90 | (4) | 6.95 ± 0.01 | 1.72 | (4) |
| I519D | 2.5 ± 0.4 ** | 97 ± 1 ** | 0.98 | (4) | a | ||
| I519F | 8.1 ± 0.1 | 82 ± 2 | 0.94 | (3) | 6.96 ± 0.02 | 1.60 | (3) |
| I519V | 11.7 ± 0.3 | 77 ± 1 | 0.84 | (3) | 6.95 ± 0.02 | 1.76 | (4) |
| A524D | 3.5 ± 0.2 ** | 90 ± 1 ** | 0.95 | (7) | 6.86 ± 0.02 ** | 1.68 | (7) |
| A524E | 2.8 ± 0.1 ** | 95 ± 1 ** | 1.00 | (7) | 7.01 ± 0.02 ** | 1.36 | (5) |
| A524K | 1.5 ± 0.1 ** | 94 ± 1 ** | 0.81 | (4) | 7.82 ± 0.04 ** | 1.01 | (4) |
| K531A | 16.4 ± 0.5 ** | 81 ± 1 | 0.95 | (4) | 7.00 ± 0.01 ** | 1.51 | (4) |
| L774A | 7.9 ± 0.7 * | 84 ± 1 * | 0.90 | (7) | 6.96 ± 0.02 | 1.52 | (3) |
| L777A | 1.1 ± 0.1 ** | 99 ± 1 ** | 0.94 | (7) | 9.13 ± 0.05 ** | 0.83 | (4–7) |
| L777C | 1.7 ± 0.1 ** | 97 ± 1 ** | 0.84 | (7) | 7.77 ± 0.05 ** | 0.91 | (3) |
| L777F | 0.9 ± 0.1 ** | 96 ± 1 ** | 0.85 | (3) | 9.18 ± 0.12 ** | 0.54 | (4) |
| L777S | 0.9 ± 0.1 ** | 98 ± 2 ** | 0.82 | (3) | 9.57 ± 0.07 ** | 0.60 | (3) |
| K778A | 15.9 ± 0.5 * | 79 ± 1 * | 0.89 | (4) | 6.90 ± 0.01 | 1.69 | (4) |
| E781A | 2.4 ± 0.1 ** | 92 ± 1 ** | 1.10 | (9) | 7.20 ± 0.06 ** | 1.22 | (8) |
| E781D | 7.4 ± 0.3 ** | 87 ± 1 ** | 0.90 | (7) | 7.14 ± 0.03 ** | 1.62 | (4) |
| E781Q | 1.3 ± 0.1 ** | 97 ± 1 ** | 1.00 | (8) | 8.18 ± 0.05 ** | 0.85 | (4) |
| NR2A mutant | |||||||
| I514A | 8.9 ± 0.4 | 81 ± 1 | 0.83 | (4) | 6.97 ± 0.02 | 1.60 | (4) |
| I514D | 16.9 ± 0.2 ** | 87 ± 1 ** | 0.93 | (4) | N.D.# | ||
| V526A | 8.4 ± 0.2 ** | 86 ± 1 ** | 0.98 | (4) | 7.02 ± 0.03 ** | 1.64 | (9) |
| V526K | 2.0 ± 0.2 ** | 94 ± 1 ** | 0.83 | (10) | 7.63 ± 0.04 ** | 1.47 | (8) |
| L777A | 2.0 ± 0.1 ** | 91 ± 1 ** | 0.90 | (2) | 7.19 ± 0.04* | 1.26 | (3) |
| L780A | 1.2 ± 0.1 ** | 97 ± 2 ** | 0.97 | (6) | 9.16 ± 0.10** | 0.81 | (4–8) |
| L780C | 1.2 ± 0.2 ** | 95 ± 1 ** | 0.85 | (6) | 8.16 ± 0.05** | 0.77 | (5) |
| L780S | 1.1 ± 0.2 ** | 96 ± 1 ** | 0.77 | (3) | 8.88 ± 0.01** | 0.59 | (3) |
| L780F | N.D. | 9.08 ± 0.03** | 0.97 | (4) | |||
| Q781A | 12.0 ± 0.9 | 81 ± 1 | 0.90 | (4) | 6.94 ± 0.02 | 1.67 | (3) |
| NR1 mut./NR2A mut. | |||||||
| I519A/I514A | 8.4 ± 0.5 | 88 ± 1 ** | 0.91 | (4) | 7.07 ± 0.03** | 1.46 | (4) |
| N521Y/E516Y | 10.5 ± 0.4 | 82 ± 1 | 1.00 | (4) | N.D. | ||
| E522Q/G784E | 15.7 ± 0.8 * | 71 ± 1 ** | 0.85 | (4) | 6.82 ± 0.01** | 1.95 | (5) |
| A524E/V526K-D776N | 1.6 ± 0.1 ** | 97 ± 2 ** | 0.91 | (4) | N.D. | ||
| S773N/S519E | 11.3 ± 1.3 | 77 ± 2 | 0.99 | (3) | 6.95 ± 0.02 | 1.58 | (2) |
| L774A/L777A | 1.5 ± 0.1 ** | 93 ± 1 ** | 0.98 | (4) | 8.04 ± 0.06** | 0.72 | (3) |
a
NR1-I519D/NR2Awt receptors displayed a bell-shaped pH dose-response curve (see Results) ; N.D.: not determined; N.D.
#
not determined because of a strong decrease in glutamate affinity (see Methods and Fig. S5);
*
p < 0.05 ;
**
p < 0.01 compared with wild-type with Student’s t test modified for unequal variances.