Twist is the key to the gating of mechanosensitive ion channel NOMPC

  1. Jingze Duan
  2. Chen Song

  • Curated by eLife

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    eLife Assessment

    This valuable study uses steered molecular dynamics simulations to interrogate force transmission in the mechanosensitive NOMPC channel, which plays roles including soft-touch perception, auditory function, and locomotion. The finding that the ankyrin spring transmits force through torsional rather than compression forces may help understand the entire TRP channel family. The evidence is, however, considered to be still incomplete. It could be strengthened by testing how the channel responds to different twisting and compressional force magnitudes over longer simulation times to see a full gating motion, or to prove that the partial or initial motion observed relates to physiological gating. Experimental validation of reduced mechano-sensitivity through mutagenesis of proposed ankyrin/TRP domain coupling interactions would be best to enhance the manuscript.

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Abstract

NOMPC, a tethered mechanosensitive ion channel from the transient receptor potential (TRP) family, can convert mechanical stimuli to ionic electric signals to excite neuron cells. Previous investigations revealed that a pushing force exerted on the linker helix domain or the compression of NOMPC’s ankyrin repeat (AR) domain triggers channel opening. In this study, we explored the direct mechanical cause for NOMPC channel opening as well as the torsional properties of the AR domain using all-atom molecular dynamics simulations. Our results demonstrate that a torque pointing toward the extracellular side, exerted on the amphipathic TRP domain, serves as the direct cause of driving the channel open. The coupling between the compression and twisting of the AR domain ensures that both types of deformations can open the channel. Therefore, we propose a twist-to-open model, facilitated by the compression-twist coupling property of the AR domain, to further elucidate the gating mechanism of the NOMPC channel.

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  1. Version published to 10.7554/elife.102941.1 on eLife
  2. Version published to 10.7554/elife.102941 on eLife
  3. eLife

    eLife Assessment

    This valuable study uses steered molecular dynamics simulations to interrogate force transmission in the mechanosensitive NOMPC channel, which plays roles including soft-touch perception, auditory function, and locomotion. The finding that the ankyrin spring transmits force through torsional rather than compression forces may help understand the entire TRP channel family. The evidence is, however, considered to be still incomplete. It could be strengthened by testing how the channel responds to different twisting and compressional force magnitudes over longer simulation times to see a full gating motion, or to prove that the partial or initial motion observed relates to physiological gating. Experimental validation of reduced mechano-sensitivity through mutagenesis of proposed ankyrin/TRP domain coupling interactions would be best to enhance the manuscript.

  4. eLife

    Reviewer #1 (Public review):

    Summary:

    This manuscript uses molecular dynamics simulations to understand how forces felt by the intracellular domain are coupled to the opening of the mechanosensitive ion channel NOMPC. The concept is interesting - as the only clearly defined example of an ion channel that opens due to forces on a tethered domain, the mechanism by which this occurs is yet to be fully elucidated. The main finding is that twisting of the transmembrane portion of the protein - specifically via the TRP domain that is conserved within the broad family of channels- is required to open the pore. That this could be a common mechanism utilised by a wide range of channels in the family, not just mechanically gated ones, makes the result significant. It is intriguing to consider how different activating stimuli can produce a similar activating motion within this family. However, the support for the finding can be strengthened as the authors cannot yet exclude that other forces could open the channel if given longer or at different magnitudes. In addition, they do not see the full opening of the channel, only an initial dilation. Even if we accept that twist is essential for this, it may be that it is not sufficient for full opening, and other stimuli are required.

    Strengths:

    Demonstrating that rotation of the TRP domain is the essential requirement for channel opening would have significant implications for other members of this channel family.

    Weaknesses:

    The manuscript centres around 3 main computational experiments. In the first, a compression force is applied on a truncated intracellular domain and it is shown that this creates both a membrane normal (compression) and membrane parallel (twisting) force on the TRP domain. This is a point that was demonstrated in the authors' prior eLife paper - so the point here is to quantify these forces for the second experiment.

    The second experiment is the most important in the manuscript. In this, forces are applied directly to two residues on the TRP domain with either a membrane normal (compression) or membrane parallel (twisting) direction, with the magnitude and directions chosen to match that found in the first experiment. Only the twisting force is seen to widen the pore in the triplicate simulations, suggesting that twisting, but not compression can open the pore. This result is intriguing and there appears to be a significant difference between the dilation of pore with the two force directions. However, there are two caveats to this conclusion. Firstly, is the magnitude of the forces - the twist force is larger than the applied normal force to match the result of experiment 1. However, it is possible that compression could also open the pore at the same magnitude or if given longer. It may be that twist acts faster or more easily, but I feel it is not yet possible to say it is the key and exclude the possibility that compression could do something similar. I also note that when force was applied to the AR domain in experiment 1, the pore widened more quickly than with the twisting force alone, suggesting that compression is doing something to assist with opening. Given that the forces are likely to be smaller in physiological conditions it could still be critical to have both twist and compression present. As this is the central aspect of the study, I believe that examining how the channel responds to different force magnitudes could strengthen the conclusions and recommend additional simulations be done to examine this.

    The second important consideration is that the study never sees a full pore opening, but rather a widening that is less than that seen in open state structures of other TRP channels and insufficient for rapid ion currents. This is something the authors acknowledge in their prior manuscript in eLife 2021. While this may simply be due to the limited timescale of the simulations, it needs to be clearly stated as a caveat to the conclusions. Twist may be the key to getting this dilation, but we don't know if it is the key to full pore opening. To demonstrate that the observed dilation is a first step in pore opening, then a structural comparison to open-state TRP channels would be beneficial to provide evidence that this motion is along the expected pathway of channel gating.

    Experiment three considers the intracellular domain and determines the link between compression and twisting of the intracellular AR domain. In this case, the end of the domain is twisted and it is shown that the domain compresses, the converse to the similar study previously done by the authors in which compression of the domain was shown to generate torque. While some additional analysis is provided on the inter-residue links that help generate this, this is less significant than the critical second experiment.

  5. eLife

    Reviewer #2 (Public review):

    This study uses all-atom MD simulation to explore the mechanics of channel opening for the NOMPC mechanosensitive channel. Previously the authors used MD to show that external forces directed along the long axis of the protein (normal to the membrane) result in AR domain compression and channel opening. This force causes two changes to the key TRP domains adjacent to the channel gate: 1) a compressive force pushes the TRP domain along the membrane normal, while 2) a twisting torque induces a clock-wise rotation on the TRP domain helix when viewing the bottom of the channel from the cytoplasm. Here, the authors wanted to understand which of those two changes is responsible for increasing the inner pore radius, and they show that it is the torque. The simulations in Figure 2 probe this question with different forces, and we can see the pore open with parallel forces in the membrane, but not with the membrane-normal forces. I believe this result as it is reproducible, the timescales are reaching 1 microsecond, and the gate is clearly increasing diameter to about 4 Å. This seems to be the most important finding in the paper, but the impact is limited since the authors already show how forces lead to channel opening, and this is further teasing apart the forces and motions that are actually the ones that cause the opening.

  6. eLife

    Reviewer #3 (Public review):

    Summary:

    This manuscript by Duan and Song interrogates the gating mechanisms and specifically force transmission in mechanosensitive NOMPC channels using steered molecular dynamics simulations. They propose that the ankyrin spring can transmit force to the gate through torsional forces adding molecular detail to the force transduction pathways in this channel.

    3. Constant velocity or constant force
    For the SMD the authors write "and a constant velocity or constant force". It's unclear from this reviewer's perspective what is used to generate the simulation data.

    Strengths:

    Detailed, rigorous simulations coupled with a novel model for force transduction.

    Weaknesses:

    Experimental validation of reduced mechanosensitivity through mutagenesis of proposed ankyrin/TRP domain coupling interactions would greatly enhance the manuscript. I have some additional questions documented below:

    (1) The membrane-parallel torsion force can open NOMPC
    How does the TRP domain interact with the S4-S5 linker? In the original structural studies, the coordination of lipids in this region seems important for gating. In this manner does the TRP domain and S4-S5 linker combined act like an amphipathic helix as suggested first for MscL (Bavi et al., 2016 Nature Communications) and later identified in many MS channels (Kefauver et al., 2020 Nature).

    (2) Torsional forces on shorter ankyrin repeats of mammalian TRP channels
    Is it possible torsional forces applied to the shorter ankyrin repeats of mammalian TRPs may also convey force in a similar manner?

    (3) Constant velocity or constant force
    For the SMD the authors write "and a constant velocity or constant force". It's unclear from this reviewer's perspective which is used to generate the simulation data.

  7. Version published to 10.1101/2024.09.26.615110v1 on bioRxiv