3E2E

Crystal Structure of an Intermediate Complex of T7 RNAP and 7nt of RNA


X-RAY DIFFRACTION

Crystallization

Crystalization Experiments
IDMethodpHTemperatureDetails
1VAPOR DIFFUSION, SITTING DROP8285PEG 6000, MgCl2, Tris ph8.0 at 285K, VAPOR DIFFUSION, SITTING DROP
Crystal Properties
Matthews coefficientSolvent content
2.7855.72

Crystal Data

Unit Cell
Length ( Å )Angle ( ˚ )
a = 81.008α = 90
b = 81.008β = 90
c = 358.791γ = 120
Symmetry
Space GroupP 61

Diffraction

Diffraction Experiment
ID #Crystal IDScattering TypeData Collection TemperatureDetectorDetector TypeDetailsCollection DateMonochromatorProtocol
11x-rayCCDADSC QUANTUM 3152007-12-10SINGLE WAVELENGTH
Radiation Source
ID #SourceTypeWavelength ListSynchrotron SiteBeamline
1SYNCHROTRONAPS BEAMLINE 24-ID-C1.5418APS24-ID-C

Data Collection

Overall
ID #Resolution (High)Resolution (Low)Percent Possible (Observed)R Merge I (Observed)Net I Over Average Sigma (I)RedundancyNumber Reflections (All)Number Reflections (Observed)Observed Criterion Sigma (F)Observed Criterion Sigma (I)B (Isotropic) From Wilson Plot
12.95093.60.0816.427490
Highest Resolution Shell
ID #Resolution (High)Resolution (Low)Percent Possible (All)Percent Possible (Observed)R Merge I (Observed)Mean I Over Sigma (Observed)RedundancyNumber Unique Reflections (All)
12.9349.40.622.5

Refinement

Statistics
Diffraction IDStructure Solution MethodCross Validation methodResolution (High)Resolution (Low)Number Reflections (Observed)Number Reflections (R-Free)Percent Reflections (Observed)R-Factor (Observed)R-WorkR-FreeR-Free Selection DetailsMean Isotropic B
X-RAY DIFFRACTIONMOLECULAR REPLACEMENTTHROUGHOUT337.824767133098.480.242960.240440.29162RANDOM99.918
Temperature Factor Modeling
Anisotropic B[1][1]Anisotropic B[1][2]Anisotropic B[1][3]Anisotropic B[2][2]Anisotropic B[2][3]Anisotropic B[3][3]
0.820.410.82-1.23
RMS Deviations
KeyRefinement Restraint Deviation
r_dihedral_angle_2_deg24.431
r_dihedral_angle_3_deg11.638
r_dihedral_angle_4_deg9.048
r_dihedral_angle_1_deg2.93
r_angle_refined_deg1.181
r_nbtor_refined0.299
r_nbd_refined0.203
r_scangle_it0.165
r_symmetry_vdw_refined0.161
r_mcangle_it0.143
RMS Deviations
KeyRefinement Restraint Deviation
r_dihedral_angle_2_deg24.431
r_dihedral_angle_3_deg11.638
r_dihedral_angle_4_deg9.048
r_dihedral_angle_1_deg2.93
r_angle_refined_deg1.181
r_nbtor_refined0.299
r_nbd_refined0.203
r_scangle_it0.165
r_symmetry_vdw_refined0.161
r_mcangle_it0.143
r_xyhbond_nbd_refined0.138
r_symmetry_hbond_refined0.114
r_scbond_it0.102
r_chiral_restr0.097
r_mcbond_it0.085
r_bond_refined_d0.006
r_gen_planes_refined0.002
r_bond_other_d
r_angle_other_deg
r_gen_planes_other
r_nbd_other
r_nbtor_other
r_xyhbond_nbd_other
r_metal_ion_refined
r_metal_ion_other
r_symmetry_vdw_other
r_symmetry_hbond_other
r_symmetry_metal_ion_refined
r_symmetry_metal_ion_other
r_mcbond_other
r_rigid_bond_restr
r_sphericity_free
r_sphericity_bonded
Non-Hydrogen Atoms Used in Refinement
Non-Hydrogen AtomsNumber
Protein Atoms6589
Nucleic Acid Atoms1351
Solvent Atoms73
Heterogen Atoms

Software

Software
Software NamePurpose
DENZOdata reduction
SCALEPACKdata scaling
PHASERphasing
REFMACrefinement
PDB_EXTRACTdata extraction