3O6Y

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	2009-07-01
Detector	RIGAKU SATURN 944+
Wavelength(s)	1.5418
Spacegroup name	P 31 2 1
Unit cell lengths	62.598, 62.598, 123.698
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	54.230 - 2.091
R-factor	0.23582
Rwork	0.233
R-free	0.29088
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1A53 with designed and adjacent residues as alanine
RMSD bond length	0.014
RMSD bond angle	1.474
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	REFMAC (5.5.0102)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	54.233	2.160
High resolution limit [Å]	2.090	2.090
Number of reflections	15690
<I/σ(I)>	16.3	5.76
Completeness [%]	96.8	87.8
Redundancy	5	4.4

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 NaAcetate pH5.5, VAPOR DIFFUSION, HANGING DROP, temperature 298K