(A) Domain architecture of TMEM163 with its structural homolog, ZnT3, with FLAG epitope tag at the C terminus of ZnT3 (ZnT3-FLAG).
(B) 300 nM ATP-evoked currents were measured from oocytes injected with combinations of cRNAs as indicated (P2X3R 25 pg; all others 2 ng) using TEVC recording (Vh = −30 mV, n = 7–9).
(C) Expression of ZnT3-FLAG was confirmed at the expected molecular weight by western blotting (WB) of cRNA-injected oocyte lysate with anti-FLAG antibody.
(D–F) Representative traces and quantification of agonist-evoked currents in cRNA-injected oocytes. cRNAs of 100 pg of P2X3R, 500 pg of GluK2 and 2 ng of Neto2, 1 ng of GluA1, 2 ng of TMEM163, and 2 ng of control (Neto2 for P2XR and GluA1 and ZnT3 for GluK2) were injected individually. TMEM163 co-expression enhanced 300 nM ATP-evoked currents of P2X3R (D), but not 500 μM glutamate-evoked currents of the kainate receptor, GluK2/Neto2 (E), or 10 μM glutamate with 50 μM cyclothiazide-evoked currents of the AMPA receptor, GluA1 (F) (n = 6–7).
Data are mean ± SEM. Mann-Whitney U-test (D). **p < 0.01.