3NXF

Robust computational design, optimization, and structural characterization of retroaldol enzymes

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU MICROMAX-007 HF
Temperature [K]	93
Detector technology	CCD
Collection date	2009-08-31
Detector	RIGAKU SATURN 944+
Wavelength(s)	1.54
Spacegroup name	P 31 2 1
Unit cell lengths	61.916, 61.916, 121.241
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	53.610 - 2.400
R-factor	0.22672
Rwork	0.225
R-free	0.26821
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1a53 with design and adjacent residues pared to alanine
RMSD bond length	0.011
RMSD bond angle	1.375
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	REFMAC (5.2.0019)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	53.610	2.490	2.160
High resolution limit [Å]	2.090	2.400	2.090
Number of reflections	10320
<I/σ(I)>	14.9	5.92	2.07
Completeness [%]	99.4	99.7	99.9
Redundancy	3.8	3.9	3.1

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	298	Protein at 5mg/ml in 100mm NaCl, 25mm Tris pH 7.5. Crystals grew at and near 2M ammonium sulfate, 4% PEG400, 100mm Na acetate pH 5.5, VAPOR DIFFUSION, HANGING DROP, temperature 298K