6B00

Thermostabilized mutant of human carbonic anhydrase II - A65T L100H K154N L224S L240P A248T

X-RAY DIFFRACTION

Crystallization

Crystalization Experiments
ID	Method	pH	Temperature	Details
1	VAPOR DIFFUSION, HANGING DROP		278	50 mM Tris pH 8, 2.5 M ammonium sulfate
2	VAPOR DIFFUSION, HANGING DROP		278	50 mM Tris pH 8, 2.5 M ammonium sulfate
3	VAPOR DIFFUSION, HANGING DROP		278	50 mM Tris pH 8, 2.5 M ammonium sulfate
4	VAPOR DIFFUSION, HANGING DROP		278	50 mM Tris pH 8, 2.5 M ammonium sulfate
5	VAPOR DIFFUSION, HANGING DROP		278	50 mM Tris pH 8, 2.5 M ammonium sulfate

Crystal Properties
Matthews coefficient	Solvent content
1.97	37.42
1.97	37.42
1.97	37.42
1.97	37.42
1.97	37.42

Crystal Data

Unit Cell
Length ( Å )	Angle ( ˚ )
a = 47.62	α = 90
b = 43.76	β = 100.72
c = 57.99	γ = 90

Symmetry
Space Group	P 1 21 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 315r		2015-02-08	M	SINGLE WAVELENGTH
2	2	x-ray	100	CCD	ADSC QUANTUM 315r		2015-09-04	M	SINGLE WAVELENGTH
3	3	x-ray	100	CCD	ADSC QUANTUM 315r		2015-09-04	M	SINGLE WAVELENGTH
4	4	x-ray	100	CCD	ADSC QUANTUM 315r		2015-09-05	M	SINGLE WAVELENGTH
5	5	x-ray	100	CCD	ADSC QUANTUM 315r		2015-09-04	M	SINGLE WAVELENGTH

Radiation Source
ID #	Source	Type	Wavelength List	Synchrotron Site	Beamline
1	SYNCHROTRON	ALS BEAMLINE 5.0.2	1.00	ALS	5.0.2
2	SYNCHROTRON	ALS BEAMLINE 5.0.2	0.886	ALS	5.0.2
3	SYNCHROTRON	ALS BEAMLINE 5.0.2	0.886	ALS	5.0.2
4	SYNCHROTRON	ALS BEAMLINE 5.0.2	0.886	ALS	5.0.2
5	SYNCHROTRON	ALS BEAMLINE 5.0.2	0.886	ALS	5.0.2

Data Collection

Overall
ID #	Resolution (High)	Resolution (Low)	Percent Possible (Observed)	Rrim 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
2	0.9	23.5	90.9	0.044	0.99	7.32	3.3		158866

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	FREE R-VALUE	1CA2	0.9	46.788	1.33	161593	8058	93.31	0.1208	0.1196	0.1442	random selection

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	12.576
f_angle_d	1.34
f_chiral_restr	0.109
f_bond_d	0.014
f_plane_restr	0.011

Non-Hydrogen Atoms Used in Refinement
Non-Hydrogen Atoms	Number
Protein Atoms	2102
Nucleic Acid Atoms
Solvent Atoms	409
Heterogen Atoms	7

Software

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

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.