Mass: 18.015 Da / Num. of mol.: 59 / Source method: isolated from a natural source / Formula: H2O
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Experimental details
-
Experiment
Experiment
Method
Number of used crystals
X-RAY DIFFRACTION
1
NEUTRON DIFFRACTION
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Sample preparation
Crystal
Density Matthews: 2.19 Å3/Da / Density % sol: 44 %
Crystal grow
Temperature: 293 K / Method: vapor diffusion, sitting drop / pH: 7.5 Details: 12-15% PEG 4000 OR PEG 3000, 0.1M MGCL2, 0.015M BETA MERCAPTOETHANOL, 0.1M TRIS-DCL, PD 7.9, 0.4M NASCN. ALL DISSOLVED IN D2O. CRYSTAL GROWN IN A 15 + 15 MICROLITRE SITTING DROP THAT WAS ...Details: 12-15% PEG 4000 OR PEG 3000, 0.1M MGCL2, 0.015M BETA MERCAPTOETHANOL, 0.1M TRIS-DCL, PD 7.9, 0.4M NASCN. ALL DISSOLVED IN D2O. CRYSTAL GROWN IN A 15 + 15 MICROLITRE SITTING DROP THAT WAS FIRST EQUILIBRATED FOR 1 WEEK. A SMALL CRYSTAL GROWN AT 20-28% PEG WAS THEN INTRODUCED. THE DROP WAS FED WITH FRESH PROTEIN BY ADDING 3-4 MICROLITRES OF PROTEIN WITH 10 MM LACTOSE EVERY 3-4 DAYS FOR 3 MONTHS. LACTOSE WAS THEN REMOVED IN TWO STEPS, FIRST BY DIALYSING AGAINST 1M GLYCEROL FOR 1 MONTH THEN AGAINST BUFFER WITH NO GLYCEROL FOR 1 MONTH. FOR DETAILS, SEE MANZONI ET AL. (2016).
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Data collection
Diffraction
ID
Mean temperature (K)
Crystal-ID
1
100
1
2
298
1
Diffraction source
Source
Site
Beamline
ID
Wavelength (Å)
SYNCHROTRON
ESRF
BM30A
1
0.98081
NUCLEAR REACTOR
ILL
LADIIII
2
3.35-4.35
Detector
Type
ID
Detector
Date
Details
ADSC QUANTUM 315r
1
CCD
Mar 10, 2017
FOCUSINGMIRRORS
2
IMAGE PLATE
Feb 15, 2017
Radiation
ID
Monochromator
Protocol
Monochromatic (M) / Laue (L)
Scattering type
Wavelength-ID
1
DOUBLECRYSTAL
SINGLEWAVELENGTH
M
x-ray
1
2
LAUE
L
neutron
2
Radiation wavelength
ID
Wavelength (Å)
Relative weight
1
0.98081
1
2
3.35
1
3
4.35
1
Reflection
Entry-ID: 6F2Q
Resolution (Å)
Num. obs
% possible obs (%)
Redundancy (%)
CC1/2
Rmerge(I) obs
Rpim(I) all
Diffraction-ID
Net I/σ(I)
Rsym value
1.03-28
70173
99.8
14
1
0.044
0.012
1
35.5
1.8-30
9844
83.8
4.9
0.993
0.053
2
8.5
0.132
Reflection shell
Resolution (Å)
Redundancy (%)
Rmerge(I) obs
Mean I/σ(I) obs
Num. unique obs
CC1/2
Rpim(I) all
Diffraction-ID
% possible all
1.03-1.06
9.3
2.357
1
5138
0.355
0.933
1
99.2
1.8-1.9
3.4
0.194
1177
0.917
0.091
2
67.1
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Processing
Software
Name
Version
Classification
PHENIX
(1.12_2829: ???)
refinement
XDS
datareduction
LAUEGEN
datascaling
PHENIX
phasing
Refinement
SU ML: 0.11 / Cross valid method: THROUGHOUT / Method to determine structure: FOURIER SYNTHESIS / Phase error: 13.66 / Shrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Starting model: 6EYM
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