2HXA

Crystal structure of Cu(I) Azurin with the metal-binding loop sequence "CTFPGHSALM" replaced with "CSPHQGAGM", at pH3.5

X-RAY DIFFRACTION

Crystallization

Crystalization Experiments
ID	Method	pH	Temperature	Details
1	VAPOR DIFFUSION, HANGING DROP	5	293	29-31% PEG 4000, 100mM magnesium chloride, 100mM Sodium acetate. Crystal soaked in ascorbate and pH3.5 buffer following growth, VAPOR DIFFUSION, HANGING DROP, temperature 293K

Crystal Properties
Matthews coefficient	Solvent content
2.23	44.81

Crystal Data

Unit Cell
Length ( Å )	Angle ( ˚ )
a = 38.338	α = 90
b = 65.726	β = 90
c = 97.577	γ = 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	93	IMAGE PLATE	RIGAKU RAXIS IV	Qsmic "blue"	2006-03-23	M	SINGLE WAVELENGTH

Radiation Source
ID #	Source	Type	Wavelength List	Synchrotron Site	Beamline
1	ROTATING ANODE	RIGAKU	1.542

Data Collection

Overall
ID #	Resolution (High)	Resolution (Low)	Percent Possible (Observed)	R Merge I (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	2.2	35.56	99.9	0.081	0.081	21	6.2		12989			29.3

Highest Resolution Shell
ID #	Resolution (High)	Resolution (Low)	Percent Possible (All)	Percent Possible (Observed)	R Merge I (Observed)	R-Sym I (Observed)	Mean I Over Sigma (Observed)	Redundancy	Number Unique Reflections (All)
	2.2	2.32	99.9		0.343	0.343	5.4	5.8	1849

Refinement

Statistics
Diffraction ID	Structure Solution Method	Cross Validation method	Resolution (High)	Resolution (Low)	Number Reflections (All)	Number Reflections (Observed)	Number Reflections (R-Free)	Percent Reflections (Observed)	R-Factor (All)	R-Factor (Observed)	R-Work	R-Free	R-Free Selection Details	Mean Isotropic B
X-RAY DIFFRACTION	Direct use of Oxidised structure in same crystal form	THROUGHOUT	2.21	35.56	12950	12950	649	99.6	0.202	0.202	0.199	0.267	RANDOM	19.472

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]
0.19			-1.74		1.55

RMS Deviations
Key	Refinement Restraint Deviation
r_dihedral_angle_2_deg	32.509
r_dihedral_angle_3_deg	13.927
r_dihedral_angle_1_deg	7.008
r_dihedral_angle_4_deg	6.372
r_scangle_it	2.088
r_scbond_it	1.422
r_angle_refined_deg	1.289
r_mcangle_it	1.016
r_mcbond_it	0.613
r_nbtor_refined	0.29

RMS Deviations
Key	Refinement Restraint Deviation
r_dihedral_angle_2_deg	32.509
r_dihedral_angle_3_deg	13.927
r_dihedral_angle_1_deg	7.008
r_dihedral_angle_4_deg	6.372
r_scangle_it	2.088
r_scbond_it	1.422
r_angle_refined_deg	1.289
r_mcangle_it	1.016
r_mcbond_it	0.613
r_nbtor_refined	0.29
r_nbd_refined	0.185
r_symmetry_vdw_refined	0.153
r_xyhbond_nbd_refined	0.111
r_symmetry_hbond_refined	0.103
r_chiral_restr	0.087
r_bond_refined_d	0.011
r_gen_planes_refined	0.004

Non-Hydrogen Atoms Used in Refinement
Non-Hydrogen Atoms	Number
Protein Atoms	1896
Nucleic Acid Atoms
Solvent Atoms	166
Heterogen Atoms	2

Software

Software
Software Name	Purpose
SCALA	data scaling
REFMAC	refinement
PDB_EXTRACT	data extraction
CrystalClear	data collection
MOSFLM	data reduction
CCP4	data scaling

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.