5A29

Family 2 Pectate Lyase from Vibrio vulnificus

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SSRL BEAMLINE BL12-2
Synchrotron site	SSRL
Beamline	BL12-2
Temperature [K]	100
Detector technology	CCD
Collection date	2013-04-22
Detector	ADSC QUANTUM 315r
Spacegroup name	P 65
Unit cell lengths	141.102, 141.102, 72.426
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	122.200 - 1.900
R-factor	0.1579
Rwork	0.156
R-free	0.19678
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2v8j
RMSD bond length	0.020
RMSD bond angle	1.907
Data reduction software	XDS
Data scaling software	SCALA
Phasing software	PHASER
Refinement software	REFMAC (5.8.0073)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	122.000	1.950
High resolution limit [Å]	1.900	1.900
Rmerge	0.100	0.420
Number of reflections	64624
<I/σ(I)>	29.1	4.6
Completeness [%]	99.8	100
Redundancy	4.8	5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7	292	CRYSTALS OF VVPL2 DEVELOPED VIA HANGING DROP VAPOUR DIFFUSION METHOD AT A PROTEIN CONCENTRATION OF 15 MG/ML BY MIXING EQUAL AMOUNTS OF THE PROTEIN SOLUTION WITH A MOTHER LIQUOR CONSISTING OF 16% (W/V) POLYETHYLENE GLYCOL 3,350, 0.14 M NA/K TARTRATE, AND 0.1 M HEPES (PH 7.0) AT 19 DEGREES CELSIUS. CRYSTALS WERE CRYOPROTECTED BY BRIEF CRYSTAL IMMERSION INTO A SOLUTION OF THE RESERVOIR SOLUTION SUPPLEMENTED WITH 25% ETHYLENE GLYCOL, AND SUBSEQUENTLY FROZEN IN A LIQUID NITROGEN STREAM PRIOR TO DIFFRACTION EXPERIMENTS.