5A6V

Open and closed conformations and protonation states of Candida antarctica Lipase B: Xenon complex

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	PETRA III, EMBL c/o DESY BEAMLINE P13 (MX1)
Synchrotron site	PETRA III, EMBL c/o DESY
Beamline	P13 (MX1)
Temperature [K]	100
Detector technology	PIXEL
Collection date	2013-03-25
Detector	DECTRIS PILATUS 6M
Spacegroup name	P 1
Unit cell lengths	39.656, 48.890, 71.680
Unit cell angles	87.80, 98.07, 108.17

Refinement procedure

Resolution	70.970 - 2.280
R-factor	0.14226
Rwork	0.139
R-free	0.19480
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1tca
RMSD bond length	0.014
RMSD bond angle	1.416
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0107)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	70.960	2.360
High resolution limit [Å]	2.270	2.270
Rmerge	0.060	0.340
Number of reflections	21330
<I/σ(I)>	29.5	7.2
Completeness [%]	92.7	70.5
Redundancy	6.2	4.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	4.8	293	CANDIDA ANTARCTICA LIPASE B (CALB) WAS PURCHASED BY HAMPTON RESEARCH AND CRYSTALLIZED WITHOUT FURTHER PURIFICATION. CRYSTALLIZATION TRIALS WERE PERFORMED AT 293 K USING THE HANGING-DROP METHOD USING A QUIAGEN EASYXTAL 15-WELL PLATE. 1 UL OF A 15 MG/ML CALB SOLUTION IN 20MM NA(CH3COO) PH = 4.8 WAS DILUTED WITH 1 UL OF THE PRECIPITANT SOLUTION, MADE OF 200MM NA(CH3COO) PH = 4.8, 20% (W/V) PEG4000, AND 10-13% (V/V) 2-PROPANOL. THE DROP WAS EQUILIBRATED BY VAPOR DIFFUSION AGAINST 500 ML OF THE PRECIPITANT SOLUTION. PROTEIN CRYSTALS OF NATIVE CALB APPEARED WITHIN ONE WEEK AND GREW TO A SIZE OF 0.2 X 0.4 X 0.5 MM3 IN THREE WEEKS