6FMB

Crystal structure of the BEC1054 RNase-like effector from the fungal pathogen Blumeria graminis

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	DIAMOND BEAMLINE I04
Synchrotron site	Diamond
Beamline	I04
Temperature [K]	80
Detector technology	PIXEL
Collection date	2015-01-01
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	0.92, 0.95
Spacegroup name	P 62 2 2
Unit cell lengths	60.520, 60.520, 78.280
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	52.410 - 1.300
R-factor	0.14536
Rwork	0.144
R-free	0.17645
Structure solution method	SAD
RMSD bond length	0.011
RMSD bond angle	1.568
Data reduction software	xia2
Data scaling software	xia2
Phasing software	SHARP
Refinement software	REFMAC (5.8.0103)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	52.510	1.340
High resolution limit [Å]	1.300	1.300
Number of reflections	21286	1483
<I/σ(I)>	23.4	5
Completeness [%]	99.5
Redundancy	6
CC(1/2)	0.998	0.996

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5	293	Purified CSEP0064/BEC1054 was dialysed into crystallisation buffer (10 mM Tris, 150 mM NaCl, pH 7.0) and concentrated to 10 mg/ml for crystallisation. Commercially available solution conditions for crystallisation (Hampton Research,CA, USA) were screened. The protein was combined with the mother liquor on a 1:1 ratio in 200 nl drops. Crystals obtained in 0.1 M sodium acetate buffer pH 5.0, supplemented with 30% PEG 4000, 0.4 M (NH4)2SO4 were cryoprotected with 30% glycerol and flash frozen for data collection.