Extended Data Fig. 6: The binding sites function evaluation.
a, The representative current traces of hTRPV3 mutants in the VSLD-PD binding sites in response to 2-APB (300 μM, red bar) and co-application of increasing concentrations of Trpvicin (from 1 nM to 100 μM, black bar), at ±80 mV. b, The representative current traces of hTRPV3-G573S mutants in the pore binding sites and the VSLD-PD binding sites inhibited by increasing concentrations of Trpvicin at ±80 mV. 10 μM RR was used to assess the whole amplitudes of leak currents. c, The representative current traces of hTRPV3 mutants in the pore binding sites in response to 2-APB and co-application of increasing concentrations of Trpvicin at ±80 mV. d, Per-residues binding energy calculated by MM-GBSA. Data are presented as mean values +/− SD of triplicates. e, The hTRPV3 mutants function similarly to the hTRPV3-WT in response to 300 μM 2-APB.The current density at −80 mV of hTRPV3 mutant channels activated by 300 μM 2-APB, compared with hTRPV3-WT. One-way ANOVA followed by Bonferroni post-tests (WT, n = 6; A556V, n = 4; A560T, n = 4; F597Y, n = 4; F601A, n = 4; T660A, n = 5; T665A, n = 5; F666A, n = 5; F666Y, n = 5). Data are presented as mean values + /- SEM. WT, wild type. f, The double mutation variants function similarly to the hTRPV3-G573S. The current density of leak currents at −80 mV of hTRPV3 double mutation variants, compared with hTRPV3-G573S. One-way ANOVA followed by Bonferroni post-tests (hTRPV3-G573S, n = 5; G573S-A556V, n = 4, G573S-A560T, n = 4; G573S-T665A, n = 4; G573S-F666A, n = 5; G573S-F666Y, n = 4). Data are presented as mean values + /- SEM; n.s., not significant.
