5FAX

Structure of subtilase SubHal from Bacillus halmapalus

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID14-2
Synchrotron site	ESRF
Beamline	ID14-2
Temperature [K]	100
Detector technology	CCD
Collection date	2002-03-01
Detector	ADSC QUANTUM 4
Wavelength(s)	0.933
Spacegroup name	P 1 21 1
Unit cell lengths	45.224, 99.935, 96.749
Unit cell angles	90.00, 103.52, 90.00

Refinement procedure

Resolution	30.000 - 2.000
R-factor	0.21522
Rwork	0.215
R-free	0.30300
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	Complex of SubHal with CI2A inhibitor PDB ID 5FBZ
RMSD bond length	0.017
RMSD bond angle	1.678
Data reduction software	HKL-2000
Data scaling software	SCALEPACK (1.97.2)
Phasing software	MOLREP
Refinement software	REFMAC (5.1.24)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	30.000	2.050
High resolution limit [Å]	2.000	2.000
Rmerge	0.101	0.268
Number of reflections	41891
<I/σ(I)>	6	2.4
Completeness [%]	74.0	49.1
Redundancy	4.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	291	Protein concentration 24 mg ml-1 in 50 mM sodium cacodylate buffer, pH 6.5, 2 mM CaCl2 and 100 mM NaCl. The enzyme was crystallised by hanging drop vapour diffusion in drops containing 1 microliter of protein and 1 microliter of reservoir solution over 0.5 ml of reservoir: Hampton Screen 2 no. 23 (10% dioxane, 0.1 M 2-(N-Morpholino)-ethanesulphonic acid (MES) buffer, pH 6.5, 1.6 M ammonium sulphate). Clusters of elongated plates appeared after several days.