Mass: 18.015 Da / Num. of mol.: 297 / Source method: isolated from a natural source / Formula: H2O
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Experimental details
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Experiment
Experiment
Method: X-RAY DIFFRACTION
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Sample preparation
Crystal
Density Matthews: 2.45 Å3/Da / Density % sol: 50 %
Crystal grow
Temperature: 298 K / Method: batch mode / pH: 4.5 Details: For the crystals grown in the presence of freshly made 3-HP, the batch reservoir consisted of 280 mM KCl, 46% PEP 426, and 140 mM sodium acetate (pH 4.5). The protein solution consisted of 6. ...Details: For the crystals grown in the presence of freshly made 3-HP, the batch reservoir consisted of 280 mM KCl, 46% PEP 426, and 140 mM sodium acetate (pH 4.5). The protein solution consisted of 6.0 mg/ml MR purified as described above, 50mM HEPES buffer (pH 7.5), and 3.3 mM MgCl2. 20mM of 3-HP was added to the protein solution before being mixed with the batch reservoir. The protein solution and reservoir solution were gently mixed in a 1:1 ratio. The mixture was pipetted into a lightly greased well to prevent the crystals from adhering to the well bottom and sealed with a glass coverslip lined with mineral oil. The spontaneous crystals grew for 17 hours, resulting in hexagonal prism and octahedral shaped crystals, and were looped directly from the wells to be flash-cooled in liquid nitrogen for storage.
Type: MARMOSAIC 300 mm CCD / Detector: CCD / Date: Aug 1, 2014
Radiation
Monochromator: Si(111) / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelength
Wavelength: 0.979 Å / Relative weight: 1
Reflection
Resolution: 1.65→50 Å / Num. obs: 49040 / % possible obs: 99.6 % / Redundancy: 13 % / Biso Wilson estimate: 17 Å2 / Rmerge(I) obs: 0.082 / Net I/σ(I): 13.2
Reflection shell
Resolution: 1.65→1.68 Å / Redundancy: 8.4 % / Rmerge(I) obs: 0.437 / Mean I/σ(I) obs: 3.2 / % possible all: 94
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Processing
Software
Name
Version
Classification
REFMAC
5.7.0029
refinement
DENZO
datareduction
SCALEPACK
datascaling
PHASER
phasing
Refinement
Method to determine structure: MOLECULAR REPLACEMENT / Resolution: 1.65→40 Å / Cor.coef. Fo:Fc: 0.972 / Cor.coef. Fo:Fc free: 0.96 / SU B: 1.255 / SU ML: 0.044 / Cross valid method: THROUGHOUT / ESU R: 0.072 / ESU R Free: 0.076 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN USED IF PRESENT IN THE INPUT
Rfactor
Num. reflection
% reflection
Selection details
Rfree
0.17677
2482
5.1 %
RANDOM
Rwork
0.14304
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obs
0.14471
46554
99.51 %
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Solvent computation
Ion probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å / Solvent model: BABINET MODEL WITH MASK