4YDN

High resolution crystal structure of human transthyretin bound to ligand and conjugates of 4-(5-(3,5-dichloro-4-hydroxyphenyl)-1,3,4-oxadiazol-2-yl)phenyl fluorosulfate

X-RAY DIFFRACTION

Crystallization

Crystalization Experiments
ID	Method	pH	Temperature	Details
1	VAPOR DIFFUSION, SITTING DROP	5.5	298	The wt-TTR was concentrated to 4 mg/ml in 10 mM NaPi, 100 mM KCl, at pH 7.6 and co-crystallized at room temperature with inhibitors using the vapor-diffusion sitting drop method, crystals were grown from 1.395 M sodium citrate, 3.5% v/v glycerol at ph 5.5. The crystals were frozen using a cryo-protectant solution of 1.395 m sodium citrate, ph 5.5, containing 10% v/v glycerol, vapor diffusion, sitting drop, temperature 298.0

Crystal Properties
Matthews coefficient	Solvent content
2.33	42.77

Crystal Data

Unit Cell
Length ( Å )	Angle ( ˚ )
a = 43.093	α = 90
b = 85.522	β = 90
c = 64.65	γ = 90

Symmetry
Space Group	P 21 21 2

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	PSI PILATUS 6M		2013-11-22	M	SINGLE WAVELENGTH

Radiation Source
ID #	Source	Type	Wavelength List	Synchrotron Site	Beamline
1	SYNCHROTRON	SSRL BEAMLINE BL11-1	0.97945	SSRL	BL11-1

Data Collection

Overall
ID #	Resolution (High)	Resolution (Low)	Percent Possible (Observed)	R Sym I (Observed)	Rrim I (All)	Rpim I (All)	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	1.349	38.484	99.5	0.028	0.031	0.012	28.4	6.4	53165	53165			14.6

Highest Resolution Shell
ID #	Resolution (High)	Resolution (Low)	Percent Possible (All)	Percent Possible (Observed)	R Merge I (Observed)	R-Sym I (Observed)	Rpim I (All)	Mean I Over Sigma (Observed)	Redundancy	Number Unique Reflections (All)
1	1.35	1.42	98		0.449	0.449	0.194	1.6	6.2	7514

Refinement

Statistics
Diffraction ID	Structure Solution Method	Cross Validation method	Starting model	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	MOLECULAR REPLACEMENT	THROUGHOUT	2QGB	1.35	38.484	50399	2693	99.4	0.1591	0.1574	0.1951	RANDOM	21.614

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]
0.13			-0.32		0.19

RMS Deviations
Key	Refinement Restraint Deviation
r_dihedral_angle_2_deg	30.791
r_sphericity_free	20.602
r_dihedral_angle_4_deg	15.481
r_dihedral_angle_3_deg	13.769
r_sphericity_bonded	8.128
r_dihedral_angle_1_deg	5.935
r_rigid_bond_restr	3.012
r_mcangle_it	2.417
r_mcbond_it	2.092
r_mcbond_other	2.013

RMS Deviations
Key	Refinement Restraint Deviation
r_dihedral_angle_2_deg	30.791
r_sphericity_free	20.602
r_dihedral_angle_4_deg	15.481
r_dihedral_angle_3_deg	13.769
r_sphericity_bonded	8.128
r_dihedral_angle_1_deg	5.935
r_rigid_bond_restr	3.012
r_mcangle_it	2.417
r_mcbond_it	2.092
r_mcbond_other	2.013
r_angle_refined_deg	1.694
r_angle_other_deg	1.361
r_chiral_restr	0.082
r_bond_refined_d	0.015
r_gen_planes_refined	0.009
r_gen_planes_other	0.006
r_bond_other_d	0.002

Non-Hydrogen Atoms Used in Refinement
Non-Hydrogen Atoms	Number
Protein Atoms	1789
Nucleic Acid Atoms
Solvent Atoms	156
Heterogen Atoms	96

Software

Software
Software Name	Purpose
SCALA	data scaling
REFMAC	refinement
PDB_EXTRACT	data extraction
XDS	data scaling
XDS	data reduction

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.