5M3C

Structure of the hybrid domain (GGDEF-EAL) of PA0575 from Pseudomonas aeruginosa PAO1 at 2.8 Ang. with GTP and Ca2+ bound to the active site of the GGDEF domain

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	BESSY BEAMLINE 14.3
Synchrotron site	BESSY
Beamline	14.3
Temperature [K]	100
Detector technology	CCD
Collection date	2016-06-23
Detector	RAYONIX SX-165mm
Wavelength(s)	0.894
Spacegroup name	P 21 21 2
Unit cell lengths	122.307, 133.931, 69.141
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	48.100 - 2.800
R-factor	0.2452
Rwork	0.244
R-free	0.27730
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4rnj 4iob
RMSD bond length	0.003
RMSD bond angle	0.741
Data reduction software	XDS
Data scaling software	Aimless (0.5.27)
Phasing software	PHASER (2.6.1)
Refinement software	REFMAC (5.8.0155)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	48.100	48.100	2.950
High resolution limit [Å]	2.800	8.850	2.800
Rmerge	0.147	0.030	1.342
Number of reflections	28700
<I/σ(I)>	14.6
Completeness [%]	99.9	98.7	99.7
Redundancy	8.1	5.2	8.4
CC(1/2)	0.998	0.998	0.740

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		294	Protein solution: 3 mg/ml in 20 mM Tris pH 7.6, 150 mM, CaCl2 2,5mM, GTP 0.32 mM mixing volumes 1 microL (1:1) Reservoir: PEG 4K 2%, CaCl2 0,01M, KCl 0,05 M, sodium cacodilato pH 6 0,05M Cryo-protection:PEG 4K raised to 3%, addition of 20% glycerol