6GJT

Chlamydia protein Pgp3 studied at high resolution in a new crystal form

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I04
Synchrotron site	Diamond
Beamline	I04
Temperature [K]	100
Detector technology	PIXEL
Collection date	2012-04-24
Detector	DECTRIS PILATUS3 S 6M
Wavelength(s)	0.9173
Spacegroup name	P 21 21 21
Unit cell lengths	85.410, 108.250, 207.000
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	42.620 - 1.980
R-factor	0.27407
Rwork	0.273
R-free	0.29220
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4jdm
RMSD bond length	0.019
RMSD bond angle	1.838
Data reduction software	MOSFLM
Data scaling software	SCALA
Phasing software	PHASER
Refinement software	REFMAC (5.7.0032)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	45.700	2.000
High resolution limit [Å]	1.980	1.980
Rmerge	0.200	1.875
Number of reflections	130981
<I/σ(I)>	9.7	1.8
Completeness [%]	98.1	98.3
Redundancy	11.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	293	A 7mg per ml Pgp3 stock buffered with 220mM NaCl, 50mM Tris-HCl pH 7.5 and 5mM DTT was screened for suitable crystallization conditions in sitting drop vapour diffusion format using a Mosquito robot (TTP Labtech, UK). A variety of commercially available screens such as Crystal Screen HT, Index HT (Hampton Research, USA), Morpheus and PGA (Molecular Dimensions, UK) were used in 96 well MRC crystallization plates (Molecular Dimensions, UK). All screening trials contained 400 nanolitre drops, each mixed in a 1 to 1 ratio of the protein and the screen solutions. The crystallization trials were incubated at 293K. The different leads obtained from the crystallization screening trials were optimised by determining working phase diagrams as detailed in Saridakis and Chayen (Saridakis & Chayen 2000). Each optimisation trial was also scaled up to 1 microlitre drop volume and set up manually in hanging drops (Qiagen Nextal plates) and in microbatch experiments (Chayen 1999).