4C3V

Extensive counter-ion interactions with subtilisin in aqueous medium, no Cs soak

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SRS BEAMLINE PX10.1
Synchrotron site	SRS
Beamline	PX10.1
Temperature [K]	100
Detector technology	CCD
Collection date	2005-06-01
Detector	MARMOSAIC 225 mm CCD
Spacegroup name	P 21 21 21
Unit cell lengths	51.891, 55.075, 75.311
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	33.760 - 2.260
R-factor	0.16274
Rwork	0.160
R-free	0.22581
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2wuw
RMSD bond length	0.016
RMSD bond angle	1.840
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Refinement software	REFMAC (5.7.0032)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.340
High resolution limit [Å]	2.260	2.260
Rmerge	0.070	0.240
Number of reflections	10628
<I/σ(I)>	55.61	14.3
Completeness [%]	100.0	100
Redundancy	10.1	8.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1				SUBTILISIN CARLSBERG WAS PURCHASED FROM SIGMA (PRODUCT CODE: P5380) AND USED FOR CRYSTALLIZATION WITHOUT FURTHER PURIFICATION. THE PROTEIN POWDER WAS DISSOLVED IN 330 MM NA CACODYLATE BUFFER AT PH 5.6 TO A CONCENTRATION OF 10 MG/ML. SUBTILISIN WAS CRYSTALLIZED BY THE BATCH METHOD FROM BUFFER SOLUTION SATURATED WITH NA2SO4 13% (W/V) AS PRECIPITANT1. LONG NEEDLE-LIKE CRYSTALS GREW OVER A PERIOD OF TWO WEEKS TO TYPICAL DIMENSIONS 50 X 50 X 400 UM X UM X UM.