5KNN

Evolutionary gain of alanine mischarging to non-cognate tRNAs with a G4:U69 base pair

X-RAY DIFFRACTION

Crystallization

Crystalization Experiments
ID	Method	pH	Temperature	Details
1	VAPOR DIFFUSION, SITTING DROP	7.5	289	0.1 M Hepes pH 7.5, 20% Polyethylene glycol 3350, 0.2 M MgCl2

Crystal Properties
Matthews coefficient	Solvent content
2.55	51.74

Crystal Data

Unit Cell
Length ( Å )	Angle ( ˚ )
a = 52.167	α = 90.07
b = 98.26	β = 89.95
c = 201.385	γ = 90.11

Symmetry
Space Group	P 1

Diffraction

Diffraction Experiment
ID #	Crystal ID	Scattering Type	Data Collection Temperature	Detector	Detector Type	Details	Collection Date	Monochromator	Protocol
1	1	x-ray	100	PIXEL	DECTRIS PILATUS 6M		2012-11-01	M	SINGLE WAVELENGTH

Radiation Source
ID #	Source	Type	Wavelength List	Synchrotron Site	Beamline
1	SYNCHROTRON	SSRL BEAMLINE BL11-1	1.1271	SSRL	BL11-1

Data Collection

Overall
ID #	Resolution (High)	Resolution (Low)	Percent Possible (Observed)	R Merge I (Observed)	Net I Over Average Sigma (I)	Redundancy	Number Reflections (All)	Number Reflections (Observed)	Observed Criterion Sigma (F)	Observed Criterion Sigma (I)	B (Isotropic) From Wilson Plot
1	2.68	50	99.2	0.142	5.5	6.9		100341

Highest Resolution Shell
ID #	Resolution (High)	Resolution (Low)	Percent Possible (All)	Percent Possible (Observed)	R Merge I (Observed)	CC (Half)	Mean I Over Sigma (Observed)	Redundancy	Number Unique Reflections (All)
1	2.68	2.73	88.7		0.408	0.894		6.8

Refinement

Statistics
Diffraction ID	Structure Solution Method	Cross Validation method	Starting model	Resolution (High)	Resolution (Low)	Number Reflections (Observed)	Number Reflections (R-Free)	Percent Reflections (Observed)	R-Factor (Observed)	R-Work	R-Free	R-Free Selection Details	Mean Isotropic B
X-RAY DIFFRACTION	MOLECULAR REPLACEMENT	THROUGHOUT	3HXU	2.68	49.13	100341	5119	93.42	0.2179	0.216	0.2542	RANDOM	55.755

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]
-10.09	5.36	-19.53	-1.42	-27.27	11.52

RMS Deviations
Key	Refinement Restraint Deviation
r_dihedral_angle_2_deg	32.608
r_dihedral_angle_4_deg	18.377
r_dihedral_angle_3_deg	17.603
r_mcangle_it	6.913
r_dihedral_angle_1_deg	6.595
r_mcbond_it	4.478
r_mcbond_other	4.478
r_angle_other_deg	3.151
r_angle_refined_deg	1.996
r_chiral_restr	0.122

RMS Deviations
Key	Refinement Restraint Deviation
r_dihedral_angle_2_deg	32.608
r_dihedral_angle_4_deg	18.377
r_dihedral_angle_3_deg	17.603
r_mcangle_it	6.913
r_dihedral_angle_1_deg	6.595
r_mcbond_it	4.478
r_mcbond_other	4.478
r_angle_other_deg	3.151
r_angle_refined_deg	1.996
r_chiral_restr	0.122
r_bond_other_d	0.02
r_bond_refined_d	0.016
r_gen_planes_other	0.016
r_gen_planes_refined	0.015

Non-Hydrogen Atoms Used in Refinement
Non-Hydrogen Atoms	Number
Protein Atoms	28360
Nucleic Acid Atoms
Solvent Atoms
Heterogen Atoms	224

Software

Software
Software Name	Purpose
HKL-2000	data collection
HKL-2000	data scaling
REFMAC	refinement
PDB_EXTRACT	data extraction

RCSB PDB Core Operations are funded by the National Science Foundation (DBI-1832184), the US Department of Energy (DE-SC0019749), and the National Cancer Institute, National Institute of Allergy and Infectious Diseases, and National Institute of General Medical Sciences of the National Institutes of Health under grant R01GM133198.