3PYK

Human Carbonic Anhydrase II as Host for Pianostool Complexes Bearing a Sulfonamide Anchor

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SEALED TUBE
Source details	OTHER
Temperature [K]	100
Detector technology	CCD
Collection date	2011-04-14
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	1.0
Spacegroup name	P 1 21 1
Unit cell lengths	42.099, 41.493, 72.382
Unit cell angles	90.00, 104.33, 90.00

Refinement procedure

Resolution	30.600 - 1.300
R-factor	0.12947
Rwork	0.128
R-free	0.16471
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2cba
RMSD bond length	0.010
RMSD bond angle	1.454
Data reduction software	MOSFLM
Data scaling software	SCALA
Phasing software	PHASER
Refinement software	REFMAC (5.5.0109)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	30.600	1.334
High resolution limit [Å]	1.300	1.300
Rmerge		0.309
Number of reflections	59007
<I/σ(I)>		2.8
Completeness [%]	96.9	91.2
Redundancy		2.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8	277	3 uL protein solution (20 mg/mL lyophilized hCAII in 50 mM Tris-sulfate, 1 mM methyl mercuric acetate) and 5 uL precipitating buffer (2.6 M ammonium sulfate, 50 mM Tris-sulfate were mixed and equilibrated against 500 uL precipitating buffer. Single crystals were transferred into a cross-linking buffer, which was prepared from 10 uL precipitating buffer and 5 uL glutaraldehyde solution (0.8 % glutaraldehyde, 4 M ammonium sulfate, 50 mM Tris-sulfate) and equilibrated for 18 h. Cross-linked crystals were transferred into soaking buffer (9 uL precipitating buffer, 0.5 uL 60 mM {(6-benzene)Ru(bispy 3)Cl}+ in DMSO) and equilibrated for 54 h, pH 8.0, VAPOR DIFFUSION, HANGING DROP, temperature 277K