3LQ0

Zymogen structure of crayfish astacin metallopeptidase

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID23-2
Synchrotron site	ESRF
Beamline	ID23-2
Temperature [K]	100
Detector technology	CCD
Collection date	2009-10-06
Detector	MAR CCD 165 mm
Wavelength(s)	0.873
Spacegroup name	P 21 21 21
Unit cell lengths	55.910, 63.370, 69.190
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	41.920 - 1.450
R-factor	0.15218
Rwork	0.152
R-free	0.18046
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1ast
RMSD bond length	0.016
RMSD bond angle	1.532
Data reduction software	XDS
Data scaling software	SCALA
Phasing software	PHASER
Refinement software	REFMAC (5.5.0046)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	46.730	1.540
High resolution limit [Å]	1.450	1.450
Rmerge	0.044	0.474
Number of reflections	44227
<I/σ(I)>	27.3	3.8
Completeness [%]	99.8	98.9
Redundancy	7.1	6.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5.6	277	For crystallization, reservoir solutions were prepared by a Tecan robot and 200-nL crystallization drops were dispensed on 96x2-well MRC plates (Innovadyne) by a Cartesian (Genomic Solutions) nanodrop robot at the High-Throughput Crystallography Platform of the Barcelona Science Park. Best crystals appeared in a Bruker steady-temperature crystal farm at 4C with protein solution (10 mg/mL in 50mM AMPSO pH9.0) and 20% PEG 8000, 0.1M (NH4)2SO4, 0.01M MgCl2, 0.05M MES pH5.6 as reservoir solution. These conditions were efficiently scaled up to the microliter range with 24-well Cryschem crystallization dishes (Hampton Research). Crystals were cryo-protected with 16% PEG 8000, 20% glycerol, 0.1M (NH4)2SO4, 0.01M MgCl2, 0.05M MES pH5.6. , VAPOR DIFFUSION, SITTING DROP, temperature 277K