6TP8

Substrate protein interactions in the limbus region of the catalytic site of Candida antarctica Lipase B

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	EMBL/DESY, HAMBURG BEAMLINE X13
Synchrotron site	EMBL/DESY, HAMBURG
Beamline	X13
Temperature [K]	100
Detector technology	PIXEL
Collection date	2013-09-22
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.824
Spacegroup name	C 2 2 21
Unit cell lengths	89.426, 156.654, 138.084
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	67.690 - 1.550
R-factor	0.1644
Rwork	0.163
R-free	0.19210
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1tca
RMSD bond length	0.006
RMSD bond angle	0.904
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	MOLREP
Refinement software	PHENIX (1.13_2998)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	67.690	1.605
High resolution limit [Å]	1.550	1.550
Rmerge	0.121	1.460
Rmeas	0.140	1.700
Rpim	0.050	0.877
Number of reflections	138806	48047
<I/σ(I)>	8.4	1
Completeness [%]	99.2	98.05
Redundancy	7.6	7.2
CC(1/2)	0.997	0.550

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	4.8	293	Crystallization trials were performed at 293 K using the hanging-drop method using a Qiagen EasyXtal 15-well plate. A 15 mg.mL CALB solution in 20mM Na(CH3COO) pH = 4.8 was incubated with a solution of paraoxon-ethyl (SIGMA catalog n. N12816) 30mM in water for one day. 1 uL of CALB solution was diluted with 1 uL of the precipitant solution, made of 200mM Na(CH3COO) pH = 4.8, 20% (w.v) PEG4000, and 15% (v.v) glyceryl tributyrate (SIGMA catalog n. T8626). The drop was equilibrated by vapor diffusion against 500 uL of the precipitant solution. Protein crystals of the CALB complex appeared within two weeks and grew to a size of 0.1 x 0.1 x 0.05 mm3 in five weeks.