Experiment: 5M7K

X-RAY DIFFRACTION

Starting Model(s)

Initial Refinement Model(s)
Type	Source	Accession Code	Details
experimental model	PDB	4CAS

Crystallization

Crystalization Experiments
ID	Method	pH	Temperature	Details
1	BATCH MODE		293	Melted monoolein was thoroughly mixed in a ratio of 3:2 (v/v) with 0.1M HEPES, pH 7.5, 0.1 % LDAO until a viscous, transparent LCP was obtained. The formed phase was then transferred into a glass vial and sponge-phase-inducing solution (1:4 ratio) was added containing 16% Jeffamine M-600, 1M HEPES pH 7.9, 0.7 M Ammonium sulphate, 2.5% 1,2,3-Heptanetriol, which swell the cubic phase to sponge phase. After phase separation overnight, the upper phase (sponge phase) was harvested. Crystals were grown using batch crystallization in the lipidic-sponge phase. Equal amount of sponge phase and protein were mixed with 2/5 (v/v) of 1.2 M Tris-sodium citrate and allowed to incubate for several weeks.

Crystal Properties
Matthews coefficient	Solvent content
3.62	65.98

Crystal Data

Unit Cell
Length ( ? )	Angle ( ? )
a = 57.9	�� = 90
b = 84.8	�� = 90
c = 384.3	�� = 90

Symmetry
Space Group	P 21 21 21

Diffraction

Diffraction Experiment
ID #	Crystal ID	Scattering Type	Data Collection Temperature	Detector	Detector Type	Details	Collection Date	Monochromator	Protocol
1	1	x-ray	293	PIXEL	CS-PAD CXI-1		2011-02-19	M	SINGLE WAVELENGTH

Radiation Source
ID #	Source	Type	Wavelength List	Synchrotron Site	Beamline
1	FREE ELECTRON LASER	SLAC LCLS BEAMLINE CXI	1.32	SLAC LCLS	CXI

Data Collection

Overall
ID #	Resolution (High)	Resolution (Low)	Percent Possible (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	3.5	58	99.4	3.5	59		25004

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	Mean Isotropic B
X-RAY DIFFRACTION	MOLECULAR REPLACEMENT	FREE R-VALUE	4CAS	3.5	55.438	1.35	24926	1294	99.98	0.254	0.2529	0.2738

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	11.846
f_angle_d	0.712
f_chiral_restr	0.04
f_plane_restr	0.004
f_bond_d	0.003

Non-Hydrogen Atoms Used in Refinement
Non-Hydrogen Atoms	Number
Protein Atoms	9076
Nucleic Acid Atoms
Solvent Atoms
Heterogen Atoms	805

Software

Software
Software Name	Purpose
PHENIX	refinement
CrystFEL	data scaling

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.