4C3U

Extensive counter-ion interactions with subtilisin in aqueous medium, Cs derivative

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	52.167, 55.226, 75.850
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	33.920 - 2.290
R-factor	0.16
Rwork	0.156
R-free	0.23431
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2wuv
RMSD bond length	0.014
RMSD bond angle	1.718
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	2.360
High resolution limit [Å]	2.280	2.280
Rmerge	0.120	0.210
Number of reflections	10346
<I/σ(I)>	10.6	6.9
Completeness [%]	99.9	99.2
Redundancy	5.8	4.6

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