Experiment: 8P8G

X-RAY DIFFRACTION

Starting Model(s)

Initial Refinement Model(s)
Type	Source	Accession Code	Details
experimental model	PDB	3U7Q

Crystallization

Crystalization Experiments
ID	Method	pH	Temperature	Details
1	VAPOR DIFFUSION, SITTING DROP	5.6	293.15	The beta-D353G/D357G MoFe-protein was crystallized anaerobically at 17.5 mg/mL under 100% N2 atmosphere. The protein was spotted as a sitting drop to 96-Well MRC 2-Drop polystyrene Crystallization Plates (SWISSCI) containing 90 uL of crystallization solution in the reservoir. Each drop contained 0.5 uL of protein sample and 0.5 uL of crystallization solution. Crystals were obtained in the crystallization solution containing 10 % w/v Polyethylene glycol 10,000; 2 % v/v 1,4-Dioxane; 100 mM tri-Sodium citrate; pH 5.6, and 1 mM polyoxotungstate [TeW6O24]6- (TEW). Sealed plates were stored inside a Coy anaerobic chamber filled with an atmosphere of N2:H2 97:3 at 20 C. Crystals were soaked in the crystallization solution supplemented with 30% v/v ethylene glycol for a few seconds before freezing in liquid nitrogen.

Crystal Properties
Matthews coefficient	Solvent content
2.74	55.06

Crystal Data

Unit Cell
Length ( ? )	Angle ( ? )
a = 167.486	�� = 90
b = 74.469	�� = 103.25
c = 208.467	�� = 90

Symmetry
Space Group	C 1 2 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 2M-F		2021-10-16	M	SINGLE WAVELENGTH

Radiation Source
ID #	Source	Type	Wavelength List	Synchrotron Site	Beamline
1	SYNCHROTRON	SLS BEAMLINE X06DA	1.00002	SLS	X06DA

Data Collection

Overall
ID #	Resolution (High)	Resolution (Low)	Percent Possible (Observed)	R Merge I (Observed)	Rrim I (All)	Rpim I (All)	CC (Half)	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	1.55	101.46	94	0.177	0.189	0.066	0.995	8.1	8.1		176043			14

Highest Resolution Shell
ID #	Resolution (High)	Resolution (Low)	Percent Possible (All)	Percent Possible (Observed)	R Merge I (Observed)	Rrim I (All)	Rpim I (All)	CC (Half)	Mean I Over Sigma (Observed)	Redundancy	Number Unique Reflections (All)
1	1.55	1.76	80.5		1.89	2.001	0.652	0.566	1.8	9.3

Refinement

Statistics
Diffraction ID	Structure Solution Method	Cross Validation method	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	Mean Isotropic B
X-RAY DIFFRACTION	MOLECULAR REPLACEMENT	FREE R-VALUE	1.55	48.32	1.33	175938	8763	48.69	0.1566	0.155	0.1872	18.62

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	15.629
f_angle_d	1.035
f_chiral_restr	0.061
f_bond_d	0.012
f_plane_restr	0.01

Non-Hydrogen Atoms Used in Refinement
Non-Hydrogen Atoms	Number
Protein Atoms	15912
Nucleic Acid Atoms
Solvent Atoms	2176
Heterogen Atoms	285

Software

Software
Software Name	Purpose
PHENIX	refinement
autoPROC	data reduction
autoPROC	data scaling
PHASER	phasing

RCSB PDB Core Operations are funded by the U.S. National Science Foundation (DBI-2321666), 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 R01GM157729.