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- EMDB-27567: Tomogram of Piezo1 vesicles -

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Basic information

Entry
Database: EMDB / ID: EMD-27567
TitleTomogram of Piezo1 vesicles
Map dataTomogram of Piezo1 vesicles
Sample
  • Complex: Piezo vesicles
Biological speciesMus (mice)
Methodelectron tomography / cryo EM
AuthorsFu Z
Funding support United States, 4 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)NIH Grant GM43949 United States
Howard Hughes Medical Institute (HHMI) United States
National Science Foundation (NSF, United States)DMR-2051681 United States
National Science Foundation (NSF, United States)DMR-1554716 United States
Citation
Journal: Proc Natl Acad Sci U S A / Year: 2022
Title: Elastic properties and shape of the Piezo dome underlying its mechanosensory function.
Authors: Christoph A Haselwandter / Yusong R Guo / Ziao Fu / Roderick MacKinnon /
Abstract: We show in the companion paper that the free membrane shape of lipid bilayer vesicles containing the mechanosensitive ion channel Piezo can be predicted, with no free parameters, from membrane ...We show in the companion paper that the free membrane shape of lipid bilayer vesicles containing the mechanosensitive ion channel Piezo can be predicted, with no free parameters, from membrane elasticity theory together with measurements of the protein geometry and vesicle size [C. A. Haselwandter, Y. R. Guo, Z. Fu, R. MacKinnon, , 10.1073/pnas.2208027119 (2022)]. Here we use these results to determine the force that the Piezo dome exerts on the free membrane and hence, that the free membrane exerts on the Piezo dome, for a range of vesicle sizes. From vesicle shape measurements alone, we thus obtain a force-distortion relationship for the Piezo dome, from which we deduce the Piezo dome's intrinsic radius of curvature, [Formula: see text] nm, and bending stiffness, [Formula: see text], in freestanding lipid bilayer membranes mimicking cell membranes. Applying these estimates to a spherical cap model of Piezo embedded in a lipid bilayer, we suggest that Piezo's intrinsic curvature, surrounding membrane footprint, small stiffness, and large area are the key properties of Piezo that give rise to low-threshold, high-sensitivity mechanical gating.
#1: Journal: Proc.Natl.Acad.Sci.USA / Year: 2022
Title: Quantitative prediction and measurement of Piezo's membrane footprint
Authors: Haselwandter CA / Guo YR / Fu Z / MacKinnon R
History
DepositionJul 11, 2022-
Header (metadata) releaseOct 5, 2022-
Map releaseOct 5, 2022-
UpdateOct 12, 2022-
Current statusOct 12, 2022Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_27567.png

Downloads & links

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Map

FileDownload / File: emd_27567.map.gz / Format: CCP4 / Size: 19.6 GB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationTomogram of Piezo1 vesicles
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
5.2 Å/pix.
x 482 pix.
= 2506.4 Å		5.2 Å/pix.
x 2881 pix.
= 14981.199 Å		5.2 Å/pix.
x 3780 pix.
= 19656. Å

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

generated in cubic-lattice coordinate

Voxel sizeX=Y=Z: 5.2 Å
Density

Histogram

Histogram (log scale)
Minimum - Maximum-0.13044275 - 0.2275203
Average (Standard dev.)0.04173477 (±0.012403254)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin00-185
Dimensions28813780482
Spacing37802881482
CellA: 19656.0 Å / B: 14981.199 Å / C: 2506.4 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : Piezo vesicles

EntireName: Piezo vesicles
Components
  • Complex: Piezo vesicles

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Supramolecule #1: Piezo vesicles

SupramoleculeName: Piezo vesicles / type: complex / Chimera: Yes / ID: 1 / Parent: 0
Source (natural)Organism: Mus (mice)
Recombinant expressionOrganism: Homo sapiens (human)

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Experimental details

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Structure determination

Methodcryo EM
Processingelectron tomography
Aggregation stateparticle

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Sample preparation

BufferpH: 8
VitrificationCryogen name: ETHANE
SectioningOther: NO SECTIONING

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average electron dose: 103.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.0 µm / Nominal defocus min: 3.0 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Final reconstructionNumber images used: 1

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