2RIO

Structure of the dual enzyme Ire1 reveals the basis for catalysis and regulation of non-conventional splicing

X-RAY DIFFRACTION

Crystallization

Crystalization Experiments
ID	Method	pH	Temperature	Details
1	VAPOR DIFFUSION, HANGING DROP	8	294	50mM Tris-Cl (pH 8.0), 200mM KOAc, 50mM SrOAc, 10mM MgCl2 and 10% PEG 8K , VAPOR DIFFUSION, HANGING DROP, temperature 294K

Crystal Properties
Matthews coefficient	Solvent content
4.29	71.3

Crystal Data

Unit Cell
Length ( Å )	Angle ( ˚ )
a = 130.307	α = 90
b = 130.307	β = 90
c = 175.011	γ = 120

Symmetry
Space Group	P 65

Diffraction

Diffraction Experiment
ID #	Crystal ID	Scattering Type	Data Collection Temperature	Detector	Detector Type	Details	Collection Date	Monochromator	Protocol
1	1	x-ray					2006-03-24	M	SINGLE WAVELENGTH

Radiation Source
ID #	Source	Type	Wavelength List	Synchrotron Site	Beamline
1	SYNCHROTRON	APS BEAMLINE 19-BM	0.97883	APS	19-BM

Data Collection

Overall
ID #	Resolution (High)	Resolution (Low)	Percent Possible (Observed)	R Sym 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	2.4	50	100	0.061	7.2	43.4		66131	1	1

Highest Resolution Shell
ID #	Resolution (High)	Resolution (Low)	Percent Possible (All)	Percent Possible (Observed)	R-Sym I (Observed)	Mean I Over Sigma (Observed)	Redundancy	Number Unique Reflections (All)
	2.4	3.11	100		0.31	8.1	6.1	6634

Refinement

Statistics
Diffraction ID	Structure Solution Method	Cross Validation method	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	SAD	THROUGHOUT	2.4	19.83	62174	3326	100	0.22443	0.22217	0.26637	RANDOM	35.687

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]
-0.05	-0.03		-0.05		0.08

RMS Deviations
Key	Refinement Restraint Deviation
r_dihedral_angle_2_deg	35.848
r_dihedral_angle_4_deg	25.585
r_dihedral_angle_3_deg	20.362
r_dihedral_angle_1_deg	9.087
r_scangle_it	5.695
r_scbond_it	3.75
r_angle_refined_deg	2.495
r_mcangle_it	2.433
r_mcbond_it	1.324
r_nbtor_refined	0.316

RMS Deviations
Key	Refinement Restraint Deviation
r_dihedral_angle_2_deg	35.848
r_dihedral_angle_4_deg	25.585
r_dihedral_angle_3_deg	20.362
r_dihedral_angle_1_deg	9.087
r_scangle_it	5.695
r_scbond_it	3.75
r_angle_refined_deg	2.495
r_mcangle_it	2.433
r_mcbond_it	1.324
r_nbtor_refined	0.316
r_symmetry_hbond_refined	0.25
r_nbd_refined	0.246
r_metal_ion_refined	0.231
r_chiral_restr	0.23
r_symmetry_vdw_refined	0.178
r_xyhbond_nbd_refined	0.133
r_bond_refined_d	0.029
r_gen_planes_refined	0.01

Non-Hydrogen Atoms Used in Refinement
Non-Hydrogen Atoms	Number
Protein Atoms	6414
Nucleic Acid Atoms
Solvent Atoms
Heterogen Atoms	58

Software

Software
Software Name	Purpose
REFMAC	refinement
HKL-2000	data collection
HKL-2000	data reduction
HKL-2000	data scaling
SHELXS	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.