Experiment: 6U9D

X-RAY DIFFRACTION

Starting Model(s)

Initial Refinement Model(s)
Type	Source	Accession Code	Details
experimental model	PDB	5FEM	BSM complex18 (PDB code 5FEM) and the RSU of Thermotoga maritima16 (PDB code 2FGC).
experimental model	PDB	2FGC	BSM complex18 (PDB code 5FEM) and the RSU of Thermotoga maritima16 (PDB code 2FGC).

Crystallization

Crystalization Experiments
ID	Method	pH	Temperature	Details
1	VAPOR DIFFUSION, HANGING DROP		291	The crystals of Saccharomyces cerevisiae AHAS complex were obtained by the hanging drop vapor diffusion method. The catalytic subunit (70.527 kDa, 70 mg/ml) and regulatory subunit (29.625 kDa, 19.8 mg/ml) freshly thawed were mixed together in a ratio of 1:2, giving a solution where the concentration of the RSU was ~ 30% in excess compared to the CSU. The cofactors, inhibitor, and DTT were added to the solution to a final concentration of 2 mM ThDP, 1 mM FAD, 10 mM MgCl2, 1 mM BSM and 5 mM DTT. The crystallization buffer consisted of 0.2 M sodium malonate pH 7 and 20% w/v PEG 3,350. The crystallization drops consisted of equal volumes (1 ul) of well solution and enzyme complex

Crystal Properties
Matthews coefficient	Solvent content
3.19	61.99

Crystal Data

Unit Cell
Length ( ? )	Angle ( ? )
a = 368.647	�� = 90
b = 230.307	�� = 94.57
c = 183.532	�� = 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	CCD	ADSC QUANTUM 210r	MIRRORS	2018-10-26	M	SINGLE WAVELENGTH

Radiation Source
ID #	Source	Type	Wavelength List	Synchrotron Site	Beamline
1	SYNCHROTRON	AUSTRALIAN SYNCHROTRON BEAMLINE MX1	0.9537	Australian Synchrotron	MX1

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	3.19	49.115	99.5	0.141	0.151	0.054	0.996	11	7.7		250942			68.75

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	3.19	3.25	96.5		0.773	0.829	0.299	0.833		7.5

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	THROUGHOUT	BSM complex18 (PDB code 5FEM) and the RSU of Thermotoga maritima16 (PDB code 2FGC).	3.194	49.11	1.34	250831	1997	99.47	0.2053	0.2049	0.2522	1997	74.3925

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	13.264
f_angle_d	0.585
f_chiral_restr	0.021
f_plane_restr	0.003
f_bond_d	0.002

Non-Hydrogen Atoms Used in Refinement
Non-Hydrogen Atoms	Number
Protein Atoms	76015
Nucleic Acid Atoms
Solvent Atoms	44
Heterogen Atoms	1593

Software

Software
Software Name	Purpose
PHENIX	refinement
XDS	data reduction
XDS	data scaling
PHENIX	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.