Piezo’s membrane footprint and its contribution to mechanosensitivity

Haselwandter, Christoph A.; MacKinnon, Roderick

doi:10.7554/elife.41968

Cited by 128 publications

(185 citation statements)

References 41 publications

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“…to mechanical stress (Haselwandter and MacKinnon, 2018). Changes in the percentage of 185 cholesterol are expected to affect the properties of the bilayer and in particular its curvature.…”

Section: The Depth Of the Piezo1 Footprint Varies With Cholesterol Comentioning

confidence: 99%

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Computational reconstruction of the complete Piezo1 structure reveals a unique footprint and specific lipid interactions

Chong

Vecchis

Hyman

et al. 2019

Preprint

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14 Piezo1 is a critical mechanical sensor in many cells. It is activated by mechanical force thus 15 allowing cells to sense the physical environment and respond to stress. Structural data have 16suggested that Piezo1 has a curved shape. Here, we use computational approaches to model, 17for the first time, the 3D structure of the full-length Piezo1 in an asymmetric membrane. A 18 number of novel insights emerge: (i) Piezo1 creates a dome in the membrane with a trilobed 19 topology that extends beyond the radius of the protein, (ii) Piezo1 changes the lipid 20 environment in its vicinity via specific interactions with cholesterol and PIP2 molecules, (iii) 21 changes in cholesterol concentration that change the membrane stiffness result in changes in 22 the depth of the dome created by Piezo1, and iv) modelling of the N-terminal region that is 23 missing from current structures modifies Piezo1 membrane footprint, suggesting the 24 importance of this region in Piezo1 function. 25

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“…to mechanical stress (Haselwandter and MacKinnon, 2018). Changes in the percentage of 185 cholesterol are expected to affect the properties of the bilayer and in particular its curvature.…”

Section: The Depth Of the Piezo1 Footprint Varies With Cholesterol Comentioning

confidence: 99%

“…Piezo1 to mechanical force (Haselwandter and MacKinnon, 2018). The sensitivity of Piezo1 72 is tuned by membrane tension and stiffness, which may in turn be influenced by varying 73 membrane lipid composition ( to stretch, and changes to membrane lipid composition (Wu et al, 2017a).…”

Section: Introduction 26mentioning

confidence: 99%

Computational reconstruction of the complete Piezo1 structure reveals a unique footprint and specific lipid interactions

Chong

Vecchis

Hyman

et al. 2019

Preprint

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“…The membrane deformation induced by a single Piezo1 channel (membrane footprint) extends well beyond the local dome and decays within tens of nanometers ( 9 ). Hence, when neighboring channels are closer than this distance, their footprints overlap with an angle smaller than 180 degrees, thus creating an additional energy penalty for membrane deformation.…”

Section: Resultsmentioning

confidence: 99%

“…In this non-conducting conformation, the arms are arranged in tri-dimensional spirals, giving Piezos a triskelion, or propeller-like, shape when viewed perpendicularly to the membrane plane and a bowl-like shape when viewed parallel to it. This curvature around the Piezo arms creates a local curvature, or dome, in the lipid bilayer, suggests the arms sense mechanical forces transmitted from lipids by sensing tension-induced flattening of the membrane ( 6, 9, 10 ).…”

Section: Introductionmentioning

confidence: 99%

A Piezo1 Open State Reveals a Multi-fenestrated Ion Permeation Pathway

Jiang

Rosario

Botello‐Smith

et al. 2020

Preprint

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19Force-sensing Piezo channels are essential to many aspects of vertebrate physiology. Activation 20 of Piezo1 is facilitated by the presence of negative membrane lipids in the inner leaflet, such as 21 phosphatidylinositol-4,5-bisphosphate (PIP2). Here, to study how Piezo1 opens, we performed 22 tension) POPC membrane (11). We also showed that the dome rapidly flattens when membrane 60 tension is gradually increased. Despite flattening of the arms, however, the pore did not open. 61Since the Piezo1 arms are anticipated to act as mechanical levers, the shorter arms in our 62 truncated model may reduce the output force (on the pore) to the input effort (arm motion). 63Another possibility for the absence of opening motions in the pore may have come from the 64 symmetric property of our simulated bilayer: indeed, electrophysiological recordings showed 65 Piezo1 remains fully closed when reconstituted in symmetrical bilayers but spontaneously opens 66 in asymmetric bilayers containing dioleoyl-sn-glycero-3-phosphatidic acid (DOPA) or 67 lysophosphatidic acid (LPA) in the inner leaflet (12, 13). DOPA and LPA differ in the number of 68 fatty acid tails but are both negatively-charged. Interestingly, the presence of negatively-charged 69 PIP2 or phosphatidylserine (PS) lipids in the inner leaflet also promote channel activation (14-16). 70We thus reasoned that adding the missing arm regions and adding negatively charged PIP2 lipids 71 in the inner leaflet will allow us to computationally capture a Piezo1 open state. 72 73 74RESULTS 75 Piezo1 clustering induces flattening of the Piezo arms 76Our previous proof-of-concept AA simulation showed that the membrane curvature, or lipid dome, 77 imposed by the resting conformation of a truncated Piezo1 takes place over 3 μs (11). To reduce 78 computational time, here we first used a 12 μs Coarse-Grained (CG) Martini simulation to enable 79 rapid lipid diffusion and dome formation while the Piezo1 backbone was kept rigid. We performed 80 these CG simulations on both a symmetrical POPC membrane (PC:PC) and an asymmetrical 81 POPC membrane containing 5% PIP2 in the inner leaflet (PC:PC/PIP2) (Figure 1a, systems I and 82 II). The CG systems were then mapped back to an AA system and simulated it for an additional 83 2 μs (Figure 1a, systems III and IV). In all MD simulations with explicit solvent, the periodic 84 boundary conditions (PBC) create an infinite lattice where the simulated system is infinitely 85

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“…Some of the C-terminal amino acid residues predicted to interact with ceramide and sphingomyelin are close to determinants of inactivation 27 , suggesting the possibility to modulate inactivation ( Figure 1e). We also analysed the dome-like membrane indentation caused by Piezo1 channel because it is thought to be a key property regulating channel gating 28,30 . Intriguingly, the depth of the dome depended on the relative proportions of ceramide and sphingomyelin (Supplementary Figure S1).…”

Section: Predicted Interaction Of Piezo1 With Sphingomyelin and Cerammentioning

confidence: 99%

Sphingomyelinase disables Piezo1 channel inactivation to enable sustained response to mechanical force

Shi

Hyman

Vecchis

et al. 2019

Preprint

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Piezo1 channels are determinants of vascular responses to fluid flow. They importantly provide sustained response to flow, but this property contrasts with the rapid inactivation that has become a hallmark of the channels in heterologous overexpression studies. Here we reveal a mechanism by which blood vessels disable inactivation to enable sustained physiological response. Creation of a molecular model of Piezo1 channel in defined lipid membranes suggested potential modulation by sphingomyelin 2 and its product ceramide. Biological relevance was indicated by the observation that exogenous sphingomyelinase enhanced Piezo1-mediated Ca 2+ entry in cultured endothelial cells. We therefore hypothesised that endogenous sphingomyelinase suppresses channel inactivation. Remarkably, in endothelium freshly-isolated from murine artery, neutral sphingomyelinase inhibitors or genetic disruption of sphingomyelin phosphodiesterase 3 (SMPD3) caused flow-and pressure-activated Piezo1 channels to become inactivating. SMPD3 retained its ability to disable inactivation in cell-free membrane patches, providing evidence for a membrane localised effect. The data suggest that inherent inactivation of Piezo1 channels is disabled by enzymatic control of lipid environment to enable physiological response to mechanical force. random order determined by the genotype of litters. Data were generated in pairs (control mice and Smpd3 fro/fro mice) and data sets compared statistically by independent t-test without assuming equal variance. Paired t-tests were used when comparing data before and after application of flow or a substance to the same membrane patch or cell. One-way ANOVA followed by Tukey posthoc test was used for comparing multiple groups. Statistical significance was considered to exist at probability (P)

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Piezo’s membrane footprint and its contribution to mechanosensitivity

Cited by 128 publications

References 41 publications

Computational reconstruction of the complete Piezo1 structure reveals a unique footprint and specific lipid interactions

Computational reconstruction of the complete Piezo1 structure reveals a unique footprint and specific lipid interactions

A Piezo1 Open State Reveals a Multi-fenestrated Ion Permeation Pathway

Sphingomyelinase disables Piezo1 channel inactivation to enable sustained response to mechanical force

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