5JEP

Human carbonic anhydrase II (T199S) complexed with benzo[d]thiazole-2-sulfonamide

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 8.2.1
Synchrotron site	ALS
Beamline	8.2.1
Temperature [K]	100
Detector technology	CCD
Collection date	2015-04-15
Detector	ADSC QUANTUM 315r
Wavelength(s)	1.1
Spacegroup name	P 1 21 1
Unit cell lengths	42.570, 41.280, 72.820
Unit cell angles	90.00, 104.75, 90.00

Refinement procedure

Resolution	70.420 - 1.190
R-factor	0.124
Rwork	0.123
R-free	0.14750
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3s73
RMSD bond length	0.010
RMSD bond angle	1.373
Data reduction software	xia2
Data scaling software	xia2
Phasing software	PHASER
Refinement software	PHENIX (dev_1839)

Data quality characteristics

	Overall
Low resolution limit [Å]	70.420
High resolution limit [Å]	1.190
Number of reflections	135839
<I/σ(I)>	13.5
Completeness [%]	88.3
Redundancy	4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		277.15	We soaked monoclinic crystals of HCAII with BTA (EVF) by carrying out the following steps: (i) We prepared soaking solutions containing 1.32 M sodium citrate, 1 mM ZnSO4, 100 mM Tris-HCl (pH 7.8), and 5 mM ligand. (ii) We combined 4 uL of soaking solution and 1-2 crystals of HCAII within a drop on the surface of a reservoir cover (EasyXtal CrystalSupport, Qiagen). (iii) To a clear plastic reservoir in a 15-reservoir plate (EasyXtal, Qiagen), we added 1 mL of soaking solution (without ligand present). We attached the reservoir cover to the reservoir and left the entire setup at 4 deg C for 1 week.