Inactivation of Mechanically Activated Piezo1 Ion Channels Is Determined by the C-Terminal Extracellular Domain and the Inner Pore Helix

Wu, Jason H.; Young, Michael; Lewis, Amanda H.; Martfeld, Ashley N.; Kalmeta, Breanna; Grandl, Jörg

doi:10.1016/j.celrep.2017.10.120

Cited by 84 publications

(103 citation statements)

References 35 publications

(54 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The inactivation of PIEZO1 is currently described as an intrinsic process in the channel gating cycle, which can be altered by introducing mutations at specific sites in the protein [23,[35][36][37]. The R2456H mutant, a xerocytosis-causing variant of human PIEZO1, displays a significant loss of inactivation with respect to wild-type (WT) PIEZO1 ( Fig.2A), as previously reported [6].…”

Section: Electrophysiological Effects Of Membrane Cholesterol Manipulsupporting

confidence: 73%

Disruption of membrane cholesterol organization impairs the concerted activity of PIEZO1 channel clusters

Ridone

Pandžić

Vassalli

et al. 2019

Preprint

View full text Add to dashboard Cite

The essential mammalian mechanosensitive channel PIEZO1 organizes in the plasma membrane into nanometric clusters which depend on the integrity of cholesterol domains to rapidly detect applied force and especially inactivate syncronously, the most commonly altered feature of PIEZO1 in pathology. ABSTRACTThe human mechanosensitive ion channel PIEZO1 is gated by membrane tension and regulates essential biological processes such as vascular development and erythrocyte volume homeostasis. Currently, little is known about PIEZO1 plasma membrane localization and organization. Using a PIEZO1-GFP fusion protein, we investigated whether cholesterol enrichment or depletion by methyl-β-Cyclodextrin (MBCD) and disruption of membrane cholesterol organization by dynasore affects PIEZO1-GFP's response to mechanical force. Electrophysiological recordings in the cell-attached configuration revealed that MBCD caused a rightward shift in the PIEZO1-GFP pressure-response curve, increased channel latency in response to mechanical stimuli and markedly slowed channel inactivation. The same effects were seen in native PIEZO1 in N2A cells. STORM super-resolution imaging revealed that, at the nano-scale, PIEZO1-GFP channels in the membrane associate as clusters sensitive to membrane manipulation. Both cluster distribution and diffusion rates were affected by treatment with MBCD (5 mM). Supplementation of polyunsaturated fatty acids appeared to sensitize the PIEZO1-GFP response to applied pressure. Together, our results indicate that PIEZO1 function is directly dependent on the membrane mechanical properties and lateral organization of membrane cholesterol domains which coordinates the concerted activity of PIEZO1 channels.

show abstract

Section: Electrophysiological Effects Of Membrane Cholesterol Manipulsupporting

confidence: 73%

Disruption of membrane cholesterol organization impairs the concerted activity of PIEZO1 channel clusters

Ridone

Pandžić

Vassalli

et al. 2019

Preprint

View full text Add to dashboard Cite

show abstract

“…The precise molecular details of Piezo1 gating are still largely unknown. The protein is also characterised by a fast inactivation phase after gating 18,19 . Recent observations proposed a functional gate possibly regulating Piezo1 inactivation 20 .…”

Section: Tension Causes Piezo1 To Flatten and Expandmentioning

confidence: 99%

Molecular principles of Piezo1 activation by increased membrane tension

Vecchis

Beech

Kalli

2019

Preprint

View full text Add to dashboard Cite

11 SUMMARY PARAGRAPH 12 13Piezo1 is a mechanosensitive channel involved in many cellular functions and responsible for 14 sensing shear-stress and pressure forces in cells1-3. Piezo1 plays a critical role in the circulatory 15 system and tissue development. Mutations on Piezo1 are linked to human diseases such as 16 lymphedema2,4 or hematological disorders such as hemolytic anaemia5 and resistance to 17 malaria6. Hypotheses for Piezo1 gating include the "force-from-lipids" principle7,8 that 18 suggests that Piezo1 senses mechanical forces through the bilayer1,9 and a direct involvement 19 of the cytoskeleton as well as the extracellular matrix in Piezo1 activation10,11. However, the 20 molecular and structural changes underpinning the Piezo1 gating mechanism and how the 21 channel senses forces in the membrane remain unknown. Here we reveal the activation 22 mechanism of Piezo1 and the structural rearrangements that occur when Piezo1 moves from a 23closed to an open state when mechanical tension is applied to the cell membrane. Our results 24show that Piezo1's curved shape is stable in a native-like model membrane without tension 25 creating a membrane indentation with a trilobed topology. Upon stretching Piezo1 adapts to 26 the stretched bilayer by flattening and expansion of its blade region. In our simulations Piezo1 27 expands up to a planar circular area of ~680 nm 2 comparable with previous structural data and 28 hypotheses12-14. Piezo1 flattening and expansion results in changes in the beam helix tilt angle. 29 These movements result in the tilting and lateral movement of the pore lining TM37 and TM38 30 helices. This leads to the opening of the channel and to the movement of lipids that occupy 31 Piezo1 pore region outside of this region, revealing for the first time the structural changes that 32 happen during Piezo1 mechanical activation. The changes in the blade region are transmitted 33 to helices TM37 and 38 via hydrophobic interactions and by interactions of neighbouring 34 2 subunits via the elbow region. The flat structure of Piezo1 identified in this study exposes the 35 C-terminal extracellular domain (CED) that in the closed state is hidden in the membrane and 36 presumably from shear stress. Our results provide new structural data for different states of 37 Piezo1 and suggest the molecular principles by which mechanical force opens the Piezo1 38 channel, thus coupling force to physiological effect via ion permeation. 39 40 41 using the protein Cα of the C-terminal extracellular domain (CED), dashed lines (lower values), and 48 of the rest of the protein (i.e. the Piezo1 blades), straight lines (higher values). (d) Projected area of 49 the Piezo1 membrane indentation on the membrane plane (Aproj, indicated by a dashed blue circle in 50 a) as a function of time. For the -40 system the timeline is the average between the two repeats. The 51 standard deviation is indicated. 52 53To investigate how mechanical forces act on the Piezo1 channel, we simulated the recent Piezo1 54 structure solved by cr...

show abstract

“…Furthermore, Piezo1 is known to regulate the mechanotransductive release of ATP into the extracellular environment . Piezo1 possesses a self‐regulatory gating mechanism at the globular C‐terminal extracellular domain, which is crucial for frequency filtering of repetitive mechanical stimuli and loading intensity . This gating mechanism of Piezo1 allows the transitions between open, closed, and inactivated stages .…”

Section: Discussionmentioning

confidence: 99%

High shear stress amplitude in combination with prolonged stimulus duration determine induction of osteoclast formation by hematopoietic progenitor cells

et al. 2020

View full text Add to dashboard Cite

To date, it is unclear how fluid dynamics stimulate mechanosensory cells to induce an osteoprotective or osteodestructive response. We investigated how murine hematopoietic progenitor cells respond to 2 minutes of dynamic fluid flow stimulation with a precisely controlled sequence of fluid shear stresses. The response was quantified by measuring extracellular adenosine triphosphate (ATP), immunocytochemistry of Piezo1, and sarcoplasmic/endoplasmic Ca2+ reticulum ATPase 2 (SERCA2), and by the ability of soluble factors produced by mechanically stimulated cells to modulate osteoclast differentiation. We rejected our initial hypothesis that peak wall shear stress rate determines the response of hematopoietic progenitor cells to dynamic fluid shear stress, as it had only a minor correlation with the abovementioned parameters. Low stimulus amplitudes corresponded to activation of Piezo1, SERCA2, low concentrations of extracellular ATP, and inhibition of osteoclastogenesis and resorption area, while high amplitudes generally corresponded to osteodestructive responses. At a given amplitude (3 Pa) and waveform (square), the duration of individual stimuli (duty cycle) showed a strong correlation with the release of ATP and osteoclast number and resorption area. Collectively, our data suggest that hematopoietic progenitor cells respond in a viscoelastic manner to loading, since a combination of high shear stress amplitude and prolonged duty cycle is needed to trigger an osteodestructive response. Plain Language Summary In case of painful joints or missing teeth, the current intervention is to replace them with an implant to keep a high‐quality lifestyle. When exercising or chewing, the cells in the bone around the implant experience mechanical loading. This loading generally supports bone formation to strengthen the bone and prevent breaking, but can also stimulate bone loss when the mechanical loading becomes too high around orthopedic and dental implants. We still do not fully understand how cells in the bone can distinguish between mechanical loading that strengthens or weakens the bone. We cultured cells derived from the bone marrow in the laboratory to test whether the bone loss response depends on (i) how fast a mechanical load is applied (rate), (ii) how intense the mechanical load is (amplitude), or (iii) how long each individual loading stimulus is applied (duration). We mimicked mechanical loading as it occurs in the body, by applying very precisely controlled flow of fluid over the cells. We found that a mechanosensitive receptor Piezo1 was activated by a low amplitude stimulus, which usually strengthens the bone. The potential inhibitor of Piezo1, namely SERCA2, was only activated by a low amplitude stimulus. This happened regardless of the rate of application. At a constant high amplitude, a longer duration of the stimulus enhanced the bone‐weakening response. Based on these results we deduce that a high loading amplitude tends to be bone weakening, and the longer this high amplitude persists, the wor...

show abstract

Inactivation of Mechanically Activated Piezo1 Ion Channels Is Determined by the C-Terminal Extracellular Domain and the Inner Pore Helix

Cited by 84 publications

References 35 publications

Disruption of membrane cholesterol organization impairs the concerted activity of PIEZO1 channel clusters

Disruption of membrane cholesterol organization impairs the concerted activity of PIEZO1 channel clusters

Molecular principles of Piezo1 activation by increased membrane tension

High shear stress amplitude in combination with prolonged stimulus duration determine induction of osteoclast formation by hematopoietic progenitor cells

Contact Info

Product

Resources

About