Mass: 26798.936 Da / Num. of mol.: 1 / Source method: obtained synthetically Details: in vitro T7 RNA polymerase with 5'cis-hammerhead ribozyme and 5'-trans VS ribozyme
Mass: 18.015 Da / Num. of mol.: 5 / Source method: isolated from a natural source / Formula: H2O
-
Experimental details
-
Experiment
Experiment
Method: X-RAY DIFFRACTION / Number of used crystals: 1
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Sample preparation
Crystal
Density Matthews: 2.36 Å3/Da / Density % sol: 47.98 %
Crystal grow
Temperature: 295 K / Method: vapor diffusion, sitting drop / pH: 7 Details: growth: 29% PEG 2000, 15 mM CaCl2, 0.2 M NH4Cl, 50 mM succinate pH 7; 0.25 mM TMP, 0.25 mM spermine, 1.5 mM MgCl2, 50 mM KCl; cryo: above with 15% w/v sucrose; 500 x 70 x 70 um crystal, pH 7. ...Details: growth: 29% PEG 2000, 15 mM CaCl2, 0.2 M NH4Cl, 50 mM succinate pH 7; 0.25 mM TMP, 0.25 mM spermine, 1.5 mM MgCl2, 50 mM KCl; cryo: above with 15% w/v sucrose; 500 x 70 x 70 um crystal, pH 7.0, VAPOR DIFFUSION, SITTING DROP, temperature 295K
Components of the solutions
ID
Name
Crystal-ID
Sol-ID
1
PEG2000
1
1
2
CaCl2
1
1
3
NH4Cl
1
1
4
MgCl2
1
1
5
KCl
1
1
6
succinate
1
1
7
spermine
1
1
8
cacodylate
1
1
9
TMP
1
1
10
PEG2000
1
2
11
CaCl2
1
2
12
NH4Cl
1
2
13
MgCl2
1
2
14
KCl
1
2
15
succinate
1
2
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Data collection
Diffraction
Mean temperature: 100 K
Diffraction source
Source: SYNCHROTRON / Site: ALS / Beamline: 8.2.2 / Wavelength: 0.95 Å
Detector
Detector: CCD / Date: Jul 10, 2005
Radiation
Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelength
Wavelength: 0.95 Å / Relative weight: 1
Reflection
Resolution: 2.89→30 Å / Num. all: 5542 / Num. obs: 5263 / % possible obs: 92.6 % / Redundancy: 5.4 % / Rmerge(I) obs: 0.124 / Net I/σ(I): 16.4
Reflection shell
Resolution: 2.89→2.95 Å / Redundancy: 2.6 % / Rmerge(I) obs: 0.39 / Mean I/σ(I) obs: 2.1 / Num. unique all: 201 / % possible all: 52.6
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Processing
Software
Name
Classification
CNS
refinement
DENZO
datareduction
SCALEPACK
datascaling
CNS
phasing
Refinement
Method to determine structure: MOLECULAR REPLACEMENT / Resolution: 2.89→30 Å / Cross valid method: THROUGHOUT / σ(F): 0 / Stereochemistry target values: Engh & Huber
Rfactor
Num. reflection
% reflection
Selection details
Rfree
0.266
553
-
random
Rwork
0.203
-
-
-
all
-
6049
-
-
obs
-
5263
87 %
-
Displacement parameters
Biso mean: 79.1 Å2
Refine analyze
Free
Obs
Luzzati coordinate error
0.5 Å
0.38 Å
Luzzati d res low
-
5 Å
Luzzati sigma a
0.53 Å
0.57 Å
Refinement step
Cycle: LAST / Resolution: 2.89→30 Å
Protein
Nucleic acid
Ligand
Solvent
Total
Num. atoms
0
1681
28
5
1714
Refine LS restraints
Refine-ID
Type
Dev ideal
X-RAY DIFFRACTION
c_bond_d
0.006
X-RAY DIFFRACTION
c_angle_deg
1.28
+
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