EMD-2290
First 3D model of wild type MjHSP16.5 at 60 degree Celsius by CryoEM.
EMD-2290
Single-particle14.0 Å
Deposition: 21/01/2013
Map released: 13/03/2013
Last modified: 24/07/2013
Concentration: 0.8
mg/mL
Buffer
pH: 7.4
Details: 10mM HEPES, 100mM NaCl.
Details: 10mM HEPES, 100mM NaCl.
Grid
Details: GIG holey grids (LifeTrust, China) were treated with a glow discharge machine (Master Plasmer)
Vitrification
Cryogen name: ETHANE
Chamber temperature: 9 K
Instrument: FEI VITROBOT MARK IV
Method: The samples were first heated to 60 degree Celsius in the boiler, then 3.5 microlitre samples were added to the grid and blotted for 1.5 s with blot force 2 at 100% humidity;
Chamber temperature: 9 K
Instrument: FEI VITROBOT MARK IV
Method: The samples were first heated to 60 degree Celsius in the boiler, then 3.5 microlitre samples were added to the grid and blotted for 1.5 s with blot force 2 at 100% humidity;
Microscope: FEI TITAN KRIOS
Illumination mode: FLOOD BEAM
Imaging mode: BRIGHT FIELD
Electron source: FIELD EMISSION GUN
Acceleration voltage: 300 kV
Nominal CS: 2.7 mm
Nominal defocus: 2.0 µm - 3.0 µm
Nominal magnification: 96000.0
Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER
Specimen holder details: Liquid nitrogen cooled
Illumination mode: FLOOD BEAM
Imaging mode: BRIGHT FIELD
Electron source: FIELD EMISSION GUN
Acceleration voltage: 300 kV
Nominal CS: 2.7 mm
Nominal defocus: 2.0 µm - 3.0 µm
Nominal magnification: 96000.0
Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER
Specimen holder details: Liquid nitrogen cooled
Temperature
Average: 85
K
Image Recording
[1]
Detector category:
CCD
Detector model: GATAN ULTRASCAN 4000 (4k x 4k)
Number of real images: 2119
Average electron dose per image: 20 e/Å2
Bits per pixel: 32.0
Details: Electron micrograph exposures were made with the automatic collection package Leginon.
Detector model: GATAN ULTRASCAN 4000 (4k x 4k)
Number of real images: 2119
Average electron dose per image: 20 e/Å2
Bits per pixel: 32.0
Details: Electron micrograph exposures were made with the automatic collection package Leginon.
Details: The particles were selected using an automatic selection program Gautomatch developed in Fei Sun lab (to be published). Octahedron symmetry were imposed during 3D reconstructing.
Final
reconstruction
Resolution: 14.0
Å
(
BY AUTHOR)
Resolution method: FSC 0.5 CUT-OFF
Number of images used: 4842
Algorithm: OTHER
Details: The final reconstructed density map was further sharpened by application of an amplitude correction algorithm in the program BFACTOR.
Resolution method: FSC 0.5 CUT-OFF
Number of images used: 4842
Algorithm: OTHER
Details: The final reconstructed density map was further sharpened by application of an amplitude correction algorithm in the program BFACTOR.
⌯ Applied Symmetry
Point group:
O
Software
[1]
Name | Version | Details |
---|---|---|
EMAN1 | - | - |
Final 2D classification
Number of classes:
193
CTF correction
Details:Each image
Format: CCP4
Data type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotation details: First model for reconstruction of wild type MjHSP16.5 at 60 degree Celsius
Details: ::::EMDATABANK.org::::EMD-2290::::
Data type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotation details: First model for reconstruction of wild type MjHSP16.5 at 60 degree Celsius
Details: ::::EMDATABANK.org::::EMD-2290::::
⬡ Geometry
X | Y | Z | |
---|---|---|---|
Dimensions | 256 | 256 | 256 |
Origin | -128 | -128 | -128 |
Spacing | 256 | 256 | 256 |
Voxel size | 0.933 Å | 0.933 Å | 0.933 Å |
Contour list
Primary | Level | Source |
---|---|---|
True | 1.73 | AUTHOR |