1LWS

Crystal structure of the intein homing endonuclease PI-SceI bound to its recognition sequence

Experimental procedure

Experimental method	MAD
Source type	SYNCHROTRON
Source details	APS BEAMLINE 14-BM-D
Synchrotron site	APS
Beamline	14-BM-D
Temperature [K]	200
Detector technology	CCD
Detector	ADSC QUANTUM 4
Wavelength(s)	0.9792, 0.9789, 0.9611
Spacegroup name	P 65 2 2
Unit cell lengths	123.000, 123.000, 211.700
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	25.000 * - 3.500
Rwork	0.287
R-free	0.31000
Structure solution method	MAD
RMSD bond length	0.011
RMSD bond angle	1.678 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	CNS
Refinement software	CNS

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	25.000 *	3.630
High resolution limit [Å]	3.500	3.500
Rmerge	0.059	0.201
Total number of observations	112701 *
Number of reflections	12539
<I/σ(I)>	18.1
Completeness [%]	98.6	96.8
Redundancy	9

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8 *	18 *	PEG 200, na hepes, calcium chloride, pH 7.5, VAPOR DIFFUSION, HANGING DROP, temperature 291K
1	VAPOR DIFFUSION, HANGING DROP	8 *	18 *	PEG 200, na hepes, calcium chloride, pH 7.5, VAPOR DIFFUSION, HANGING DROP, temperature 291K

Crystallization Reagents

ID	crystal ID	solution ID	reagent name	concentration	details
1	1	1	PEG 200
2	1	1	na hepes
3	1	1	CaCl2
4	1	2	CaCl2
5	1	2	na hepes

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	6 (mg/ml)
2	1	drop		60 (mM)
3	1	drop	EDTA	0.5 (mM)
4	1	drop		2.5 (mM)
5	1	drop	TCEP	1 (mM)
6	1	drop	Tris-HCl	10 (mM)	pH8.
7	1	reservoir	PEG200	25 (%(v/v))
8	1	reservoir		0.2 (M)
9	1	reservoir	sodium HEPES	0.1 (M)	pH7.5