5TDX

Resurrected Ancestral Hydroxynitrile Lyase from Flowering Plants

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 23-ID-B
Synchrotron site	APS
Beamline	23-ID-B
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	2014-10-30
Detector	MAR scanner 300 mm plate
Wavelength(s)	1.0332
Spacegroup name	P 43 21 2
Unit cell lengths	108.265, 108.265, 201.188
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	29.320 - 1.960
R-factor	0.1792
Rwork	0.177
R-free	0.22650
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1yb6
RMSD bond length	0.027
RMSD bond angle	2.346
Data reduction software	XDS
Data scaling software	Aimless (0.3.11)
Phasing software	PHASER (2.5.1)
Refinement software	REFMAC (5.8.0155)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	29.320	1.990
High resolution limit [Å]	1.960	1.960
Rmerge	0.097	0.680
Number of reflections	86348
<I/σ(I)>	13.8
Completeness [%]	99.3	88.3
Redundancy	8.5	5.8
CC(1/2)	0.998	0.772

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.8	293	1:1 mix of protein (in 5mM BES buffer) with mother liquor (1.1 Molar Ammonium Citrate, 50 mM bis-tris propane, pH 6.8) Crystals formed after 2-3 weeks at 20 degrees C. Crystals were cryo-protected in 5% glycerol in mother liquor for 10 seconds then flash frozen in LN2.