2Y75

The Structure of CymR (YrzC) the Global Cysteine Regulator of B. subtilis

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID14-4
Synchrotron site	ESRF
Beamline	ID14-4
Temperature [K]	100
Detector technology	CCD
Collection date	2008-10-26
Detector	ADSC QUANTUM 315r
Spacegroup name	P 21 21 21
Unit cell lengths	98.700, 103.700, 106.530
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	17.840 - 2.000
R-factor	0.1884
Rwork	0.187
R-free	0.21360
Structure solution method	SAD
Starting model (for MR)	NONE
RMSD bond length	0.010
RMSD bond angle	1.090
Data reduction software	XDS
Data scaling software	XDS
Refinement software	BUSTER (2.8.0)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	17.840	2.050
High resolution limit [Å]	2.000	2.000
Rmerge	0.060	0.740
Number of reflections	73831
<I/σ(I)>	14.42	2.03
Completeness [%]	99.0	99.6
Redundancy	3.64	3.68

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		291	OPTIMIZATION OF INITIAL HITS WAS PURSUED MANUALLY IN LINBRO PLATES WITH A HANGING DROP SETUP. THE BEST CRYSTALS WERE OBTAINED BY MIXING 1.5 UL OF NATIVE CYMR PROTEIN OR SELENOMETHIONINE LABELED PROTEIN AT 14 MG/ML WITH 1.5 UL OF THE RESERVOIR SOLUTION CONTAINING 1.6 M AMMONIUM SULPHATE AND 0.1 M TRIS-HCL PH 8.5 AT 18 C DEGREES. THE CRYSTALS APPEARED WITHIN ONE WEEK AND HAD DIMENSIONS OF UP TO 0.1 MM X 0.1 MM X 0.2 MM. A SINGLE CRYSTAL OF THE CYMR PROTEIN WAS FLASH-FROZEN IN LIQUID NITROGEN USING A MIXTURE OF 50% PARATONE AND 50% PARAFFIN OIL AS CRYOPROTECTANT.