Experiment: 4UAM

X-RAY DIFFRACTION

Crystallization

Crystalization Experiments
ID	Method	pH	Temperature	Details
1	VAPOR DIFFUSION, HANGING DROP	6.5	297	drop contained 1ul protein (25-30mg/ml in 20mm HEPES, 50mm NaCl, ph7.5) with 1uL microseed reservoir (0.2M sodium acetate, 0.2M sodium citrate, 26% PEGMME 2K). Protein solution was heat-treated at 310K for 3-5 days prior to filtration and crystallisation screening. microseeding was essential for diffraction quality crystal growth. crystal grew over 2 weeks

Crystal Properties
Matthews coefficient	Solvent content
2.97	58.53

Crystal Data

Unit Cell
Length ( Å )	Angle ( ˚ )
a = 49.99	α = 83.45
b = 75.91	β = 75.3
c = 82.36	γ = 74.01

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		CCD	ADSC QUANTUM 210r		2013-03-14	M	SINGLE WAVELENGTH
2	1	x-ray		CCD	ADSC QUANTUM 210r		2013-03-16	M	SINGLE WAVELENGTH

Radiation Source
ID #	Source	Type	Wavelength List	Synchrotron Site	Beamline
1	SYNCHROTRON	AUSTRALIAN SYNCHROTRON BEAMLINE MX1	1.283500	Australian Synchrotron	MX1
2	SYNCHROTRON	AUSTRALIAN SYNCHROTRON BEAMLINE MX1	1.7311	Australian Synchrotron	MX1

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	1.8	44.93	94.81	0.05521	18.08	3.9	381468				19.81

Highest Resolution Shell
ID #	Resolution (High)	Resolution (Low)	Percent Possible (All)	Percent Possible (Observed)	R Merge I (Observed)	Rrim I (All)	Mean I Over Sigma (Observed)	Redundancy	Number Unique Reflections (All)
1	1.8	1.85	92.26		0.4806	0.569	2.9	3.9

Refinement

Statistics
Diffraction ID	Structure Solution Method	Cross Validation method	Starting model	Resolution (High)	Resolution (Low)	Cut-off Sigma (F)	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	FREE R-VALUE	PDB ENTRY 2DOO	1.8	44.927	1.99	191266	9681	91.74	0.1625	0.1615	0.182	Random Selection	34.3635

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]

RMS Deviations
Key	Refinement Restraint Deviation
f_dihedral_angle_d	11.528
f_angle_d	0.962
f_chiral_restr	0.037
f_bond_d	0.006
f_plane_restr	0.005

Non-Hydrogen Atoms Used in Refinement
Non-Hydrogen Atoms	Number
Protein Atoms	6900
Nucleic Acid Atoms
Solvent Atoms	969
Heterogen Atoms	80

Software

Software
Software Name	Purpose
PDB_EXTRACT	data extraction
PHENIX	refinement
Coot	model building
PHASER	phasing
XSCALE	data scaling
XDS	data reduction
Blu-Ice	data collection

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.