6UWE

Crystal structure of recombinant thiocyanate dehydrogenase from Thioalkalivibrio paradoxus saturated with copper

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID29
Synchrotron site	ESRF
Beamline	ID29
Temperature [K]	100
Detector technology	PIXEL
Collection date	2017-12-10
Detector	DECTRIS PILATUS 6M
Wavelength(s)	0.96900
Spacegroup name	P 1 21 1
Unit cell lengths	90.422, 163.149, 90.631
Unit cell angles	90.00, 119.29, 90.00

Refinement procedure

Resolution	44.800 - 1.600
R-factor	0.1424
Rwork	0.141
R-free	0.16020
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	6i3q
RMSD bond length	0.009
RMSD bond angle	1.511
Data reduction software	HKL-3000
Data scaling software	SCALEPACK
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0257)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.630
High resolution limit [Å]	1.600	4.340	1.600
Rmerge	0.074	0.062	0.461
Rmeas	0.092	0.076	0.575
Rpim	0.053	0.044	0.341
Number of reflections	285581	14867	14013
<I/σ(I)>	9.3		2.1
Completeness [%]	94.4	97.1	92.6
Redundancy	2.6	2.6	2.6
CC(1/2)	0.993	0.977	0.692

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		293	10 mg/mL TcDH in 25 mM borate buffer, pH 9.5 was mixed with 0.17 M ammonium acetate, 0.085 M sodium citrate, pH 5.6, 25.5% w/v PEG 4000, 15% glycerol. To increase copper incorporation, powder of CuCl was added to drops with crystals 30-40 min prior to harvesting. Sodium cyanate solution was added to the drop with crystals to the final concertation of ~100 mM at the same time as CuCl was added.