5T0W

Crystal structure of the ancestral amino acid-binding protein AncCDT-1, a precursor of cyclohexadienyl dehydratase

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	AUSTRALIAN SYNCHROTRON BEAMLINE MX1
Synchrotron site	Australian Synchrotron
Beamline	MX1
Temperature [K]	100
Detector technology	CCD
Collection date	2015-10-29
Detector	ADSC QUANTUM 210r
Wavelength(s)	0.9501
Spacegroup name	P 21 21 21
Unit cell lengths	47.030, 68.880, 318.630
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	33.190 - 2.590
R-factor	0.25518
Rwork	0.253
R-free	0.28733
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4zv2
RMSD bond length	0.011
RMSD bond angle	1.372
Data reduction software	iMOSFLM (1.0.7)
Data scaling software	Aimless (0.1.29)
Phasing software	PHASER (2.5.2)
Refinement software	REFMAC (5.7.0032)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	33.660	2.710
High resolution limit [Å]	2.590	2.590
Rmerge	0.158	0.775
Number of reflections	33106
<I/σ(I)>	8.2	2.4
Completeness [%]	99.4	99.3
Redundancy	6.8	6.9
CC(1/2)	0.993	0.755

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8.2	291	A crystal grown in a hanging drop of 2 uL protein (18 mg/mL in 20 mM HEPES pH 7.5, 50 mM NaCl, 1 mM arginine) + 2 uL precipitant (0.2 M lithium sulfate, 0.1 M TRIS pH 8.2, 22% (w/v) PEG 3350) was improved by three rounds of serial microseeding; the crystals were crushed and serially diluted in the precipitant, and new hanging drops were prepared by mixing 2 uL protein and 2 uL microseed suspension.