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 drops that was first equilibrated for 1 week. A crystal grown at 20-28% PEG was introduced. The drop was fed with fresh protein by adding 3-4 micro litres of protein with 10 mM lactose every 3-4 days for 3 months. For details see Manzoni et al. (2016).
293
2
vapor diffusion
7.5
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 drops that was first equilibrated for 1 week. A crystal grown at 20-28% PEG was introduced. The drop was fed with fresh protein by adding 3-4 micro litres of protein with 10 mM lactose every 3-4 days for 3 months. For details see Manzoni et al. (2016).
-
Data collection
Diffraction
ID
Mean temperature (K)
Crystal-ID
1
298
1
2
298
1
3
298
1
Diffraction source
Source
Site
Beamline
ID
Wavelength (Å)
SYNCHROTRON
MAX II
I911-3
1
1
NUCLEAR REACTOR
ILL
LADIIII
2
3.35-4.35
SPALLATION SOURCE
ORNL Spallation Neutron Source
MANDI
3
2.0-4.0
Detector
Type
ID
Detector
Date
Details
MARMOSAIC 325 mm CCD
1
CCD
May 31, 2015
FOCUSINGMIRRORS
LADI III
2
IMAGE PLATE
Dec 15, 2014
ORNL ANGER CAMERA
3
AREA DETECTOR
May 15, 2015
Radiation
ID
Monochromator
Protocol
Monochromatic (M) / Laue (L)
Scattering type
Wavelength-ID
1
SI(111) DOUBLECRYSTAL
SINGLEWAVELENGTH
M
x-ray
1
2
LAUE
L
neutron
2
3
LAUE
L
neutron
3
Radiation wavelength
ID
Wavelength (Å)
Relative weight
1
1
1
2
3.35
1
3
4.35
1
4
2
1
5
4
1
Reflection
Entry-ID: 6EYM
Resolution (Å)
Num. obs
% possible obs (%)
Redundancy (%)
CC1/2
Rmerge(I) obs
Diffraction-ID
Net I/σ(I)
Rpim(I) all
1.07-40
61354
99.4
6.6
1
0.032
1
24.7
1.7-28
15485
95.8
13
0.209
2
22.2
0.052
1.7-28
13741
87.1
10
0.162
2
12.4
0.044
1.6-19
15485
81.3
4.4
0.155
3
12.2
0.066
Reflection shell
Resolution (Å)
Redundancy (%)
Rmerge(I) obs
Mean I/σ(I) obs
CC1/2
Diffraction-ID
% possible all
Num. unique obs
Rpim(I) all
1.07-1.1
0.817
2.1
0.772
1
96.8
1.7-1.8
4.9
0.334
3.9
2
87.9
2172
0.139
1.7-1.8
4.9
0.208
7
2
66.5
1490
0.076
1.6-1.66
1.7
0.207
2.4
3
54.6
1022
0.15
-
Processing
Software
Name
Version
Classification
PHENIX
(1.12_2829: ???)
refinement
LAUEGEN
datareduction
LSCALE
datascaling
SCALA
datascaling
Mantid
datareduction
XPREP
datascaling
PHENIX
phasing
Refinement
SU ML: 0.07 / Cross valid method: THROUGHOUT / Method to determine structure: FOURIER SYNTHESIS / Phase error: 10.82 / Shrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Starting model: 3ZSJ
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