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. 2021 Jun 8;10:e58388. doi: 10.7554/eLife.58388

Figure 2. Conformational changes of the transient receptor potential (TRP) and transmembrane (TM) domains during gating.

(A) Principal component analysis (PCA) of the molecular dynamics (MD) simulation trajectories (FI0, SI0, and CI0 in Supplementary file 1a). The projections on the second eigenvector can distinguish the conformations under pulling (red) or pushing (blue). (B) The overlaid extreme structures along the second eigenvector of the PCA. The most closed conformation (silver) and open conformation (cyan) showed the global changes of the TRP domain during gating: a clockwise rotation. (C) The orientation and position of the gate residue, I1554, in the most closed (silver) and open (cyan) conformations in the simulations. (D, E) The residues forming four stable hydrogen bonds between the TRP and LH domains. (F) The mean and standard deviation (SD) of the mechano-gated current of the wild-type NompC, as well as the mutants W1572A, S1421A, Q1253A, S1577A, K1244A, E1571A, D1236A, and R1581A, under negative pressure in the outside-out patch-clamp experiments (wild type: n = 13; W1572A: n = 7; S1421A: n = 6; Q1253A: n = 6; S1577A: n = 5; K1244A: n = 6; E1571A: n = 6; D1236A: n = 9; R1581A: n = 6). All of the error bars denote ± SD. Hydrogen bonds are indicated by dashed lines (D, E).

Figure 2.

Figure 2—figure supplement 1. The transmembrane (TM) pore size evolution and rotation angle evolution of the transient receptor potential (TRP) domain from principal component analysis (PCA).

Figure 2—figure supplement 1.

Figure 2—figure supplement 2. The conformational change of the transient receptor potential (TRP) domain in the steered molecular dynamics (SMD) simulations.

Figure 2—figure supplement 2.

The side and bottom views of the S6 and TRP domains before and after pulling or pushing.
Figure 2—figure supplement 3. The overlaid closed-state and open-state structures of TRPV1, obtained in lipid nanodisc.

Figure 2—figure supplement 3.

Figure 2—figure supplement 4. The schematic figure of pUAST-NOMPC_EGFP (del-miniwhite) used in the experiment.

Figure 2—figure supplement 4.

Figure 2—figure supplement 5. The mutants of residues listed in Figure 2D–F showed normal membrane targeting.

Figure 2—figure supplement 5.

Figure 2—figure supplement 6. Formation of the alternative hydrogen bonds in the mutant, as identified in the molecular dynamics (MD) simulations (from chain A in the trajectories D1236A and E1571A in Supplementary file 1a).

Figure 2—figure supplement 6.