Fig. 2: Functional relevance of the GluA3 NTD–LBD interface.
From: Architecture, dynamics and biogenesis of GluA3 AMPA glutamate receptors
a, Model of the apo-state interface, showing key interacting residues. LBD loop 2 and Arg395 in the NTD–LBD linker are also shown, both of which contribute to the interface. b, Representative overlaid current traces from outside-out patches of HEK293 cells expressing GluA3-R–TARP-γ2 (green) and GluA1–TARP-γ2 (black), elicited with 10 mM glutamate (1 ms pulses at 100 Hz). Currents are normalized to the first response (P1) in the application. c, Example peak-scaled whole-cell currents elicited by application of 10 mM l-glutamate for 200 ms at −60 mV from cells expressing GluA3-G–TARP-γ2 (green trace) and GluA3-G(K129C/E458C)–TARP-γ2 (black trace) (top). Inset: the mean ± s.d. steady-state (SS) current for GluA3-G–TARP-γ2 (n = 8 cells) and GluA3-R(K129C/E458C)–TARP-γ2 (n = 6 cells). Statistical analysis was performed using an unpaired two-tailed t-test; t = 12.16, d.f. = 12, P < 0.0001. Bottom, the mean ± s.d. desensitization time constants for GluA3-G–TARP-γ2 (n = 8 cells), GluA3-G(K129C)–TARP-γ2 (n = 7 cells), GluA3-G(E458C)–TARP-γ2 (n = 6 cells) and GluA3-G(K129C/E458C)–TARP-γ2 (n = 6 cells). Statistical analysis was performed using Welch’s analysis of variance (ANOVA; W3,8.572 = 11.01, P = 0.0010) followed by Dunnett’s multiple-comparison test; P values are indicated. d, Current amplitudes of the first five responses evoked from the outside-out patches using the protocol as described in b, expressing GluA3-G–TARP-γ2 (n = 11 patches), GluA3-G(K129C/E458C)–TARP-γ2 (n = 6 patches) and GluA3-G(Q185glyco)–TARP-γ2 (n = 7 patches) (left). Currents are normalized to the first pulse. Data are mean ± s.d. A dashed line connecting the dots is included as a visual guide. Inset: aligned, peak-scaled whole-cell current responses to 10 mM, 200 ms glutamate application. Right, the mean ± s.d. current ratio between the fifth and first pulses (P5/P1) for the same data. Statistical analysis was performed using one-way ANOVA (W(2,21) = 38.70, P < 0.0001) followed by Dunnett’s multiple-comparisons test; P values are indicated.
