6NX9

ECAII(D90T,K162T) MUTANT IN COMPLEX WITH CITRATE AT PH 7

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU MICROMAX-007 HF
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-04-12
Detector	DECTRIS EIGER R 4M
Wavelength(s)	1.5418
Spacegroup name	C 1 2 1
Unit cell lengths	152.009, 62.539, 143.239
Unit cell angles	90.00, 118.19, 90.00

Refinement procedure

Resolution	26.550 - 1.970
R-factor	0.1617
Rwork	0.160
R-free	0.22570
Structure solution method	FOURIER SYNTHESIS
RMSD bond length	0.020
RMSD bond angle	2.290
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	REFMAC
Refinement software	REFMAC (5.8.0238)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	40.000	40.000	2.000
High resolution limit [Å]	1.970	5.340	1.970
Rmerge	0.095	0.028	0.511
Rmeas	0.113	0.033	0.631
Rpim	0.061	0.018	0.365
Total number of observations	263837
Number of reflections	81136	4298	3900
<I/σ(I)>	7.1
Completeness [%]	96.6	98.3	92.1
Redundancy	3.3	3.3	2.6
CC(1/2)		0.997	0.674

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7	293	Protein, at the concentration 15 mg/ml in 50 mM HEPES buffer pH 7 and 150 mM sodium chloride was mixed with equivolume solution of precipitant that contained, 17% (w/v) PEG3350 and 0.17 M ammonium citrate pH 7. Resulting droplets were equilibrated against the precipitant. Before data collection, crystal was transferred the precipitant containing L-Asn at concentration 20 mM. Subsequently, for the data collection, crystal was briefly transferred to cryo-protecting solution, which had the same composition as precipitant (with L-Asn), except concentration of PEG3350 was increased to 35 % (v/w/) and 10% (v/v), and also contained 15% of glycerol