EMD-1087
Three-dimensional structure of the bacterial multidrug transporter EmrE shows it is an asymmetric homodimer.
EMD-1087
Electron Crystallography7.5 Å
Deposition: 09/06/2004Map released: 10/06/2004
Last modified: 02/10/2013
Concentration: 1
mg/mL
Details: 2D crystals were grown in suspension by dialysis
Details: 2D crystals were grown in suspension by dialysis
Buffer
pH: 7.5
Details: 20 mM Sodium phosphate pH7.5, 100 mM NaCl, 2mM MgCl2, 1mM EDTA, 1 mM Na azide, 10uM tetraphenylphosphonium
Details: 20 mM Sodium phosphate pH7.5, 100 mM NaCl, 2mM MgCl2, 1mM EDTA, 1 mM Na azide, 10uM tetraphenylphosphonium
Staining
Type:
NEGATIVE
Details: Crystals on carbon support were washed with 1% glucose pH 8 (aq ammonia) containing 25ug/ml bacitracin, 2 times 20ul, and blotted to dryness
Details: Crystals on carbon support were washed with 1% glucose pH 8 (aq ammonia) containing 25ug/ml bacitracin, 2 times 20ul, and blotted to dryness
Grid
Details: 300 mesh molybdenum grid
Vitrification
Cryogen name: NITROGEN
Chamber humidity: 45%
Chamber temperature: 77 K
Instrument: HOMEMADE PLUNGER
Method: Blot to dryness for 10 seconds before freezing
Details: Vitrification instrument: manual. Performed at room temperature
Chamber humidity: 45%
Chamber temperature: 77 K
Instrument: HOMEMADE PLUNGER
Method: Blot to dryness for 10 seconds before freezing
Details: Vitrification instrument: manual. Performed at room temperature
Crystal
formation
Details: 2D crystals were grown in suspension by dialysis
Microscope: FEI TECNAI F30
Illumination mode: SPOT SCAN
Imaging mode: BRIGHT FIELD
Electron source: FIELD EMISSION GUN
Acceleration voltage: 300 kV
Nominal CS: 2.3 mm
Nominal defocus: 0.2 µm - 1.6 µm
Nominal magnification: 60000.0
Calibrated magnification: 57400.0
Specimen holder model: GATAN LIQUID NITROGEN
Specimen holder details: Side entry liquid nitrogen-cooled cryo specimen holder
Alignment procedure: LEGACY (Astigmatism: objective lens astigmatism was corrected at 200,000 times magnification, Electron beam tilt params: )
Minimum tilt angle: 0
Maximum tilt angle: 46
Illumination mode: SPOT SCAN
Imaging mode: BRIGHT FIELD
Electron source: FIELD EMISSION GUN
Acceleration voltage: 300 kV
Nominal CS: 2.3 mm
Nominal defocus: 0.2 µm - 1.6 µm
Nominal magnification: 60000.0
Calibrated magnification: 57400.0
Specimen holder model: GATAN LIQUID NITROGEN
Specimen holder details: Side entry liquid nitrogen-cooled cryo specimen holder
Alignment procedure: LEGACY (Astigmatism: objective lens astigmatism was corrected at 200,000 times magnification, Electron beam tilt params: )
Minimum tilt angle: 0
Maximum tilt angle: 46
Temperature
Minimum: 77
K
Average: 77 K
Maximum: 77 K
Average: 77 K
Maximum: 77 K
Image Recording
[1]
Detector category:
FILM
Detector model: KODAK SO-163 FILM
Scanner: ZEISS SCAI
Sampling interval: 7 µm
Number of real images: 47
Average electron dose per image: 15 e/Å2
Detector model: KODAK SO-163 FILM
Scanner: ZEISS SCAI
Sampling interval: 7 µm
Number of real images: 47
Average electron dose per image: 15 e/Å2
Tilt Series
[1]
| Axis 1 | Axis 2 | |||||
|---|---|---|---|---|---|---|
| Min. | Max. | Inc. | Min. | Max. | Inc. | Rotation |
| 0 ° | 46 ° | - | - | - | - | - |
Format: CCP4
Data type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotation details: Density map of the asymmetric dimer of EmrE,a multidrug transporter from Escherichia coli
Details: ::::EMDATABANK.org::::EMD-1087::::
Data type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Annotation details: Density map of the asymmetric dimer of EmrE,a multidrug transporter from Escherichia coli
Details: ::::EMDATABANK.org::::EMD-1087::::
⬡ Geometry
| X | Y | Z | |
|---|---|---|---|
| Origin | -2 | -63 | -10 |
| Dimensions (px) | 81 | 89 | 21 |
| Dimensions (Å) | 72.10005 | 86.80005 | 200.0 |
| Voxel size (Å) | 0.48066702 | 0.578667 | 2.5 |
Contour list
| Primary | Level | Source |
|---|---|---|
| True | 59.7 | AUTHOR |