5KR3

Directed Evolution of Transaminases By Ancestral Reconstruction. Using Old Proteins for New Chemistries

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	AUSTRALIAN SYNCHROTRON BEAMLINE MX2
Synchrotron site	Australian Synchrotron
Beamline	MX2
Temperature [K]	100
Detector technology	CCD
Collection date	2014-02-18
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.95370
Spacegroup name	P 1 21 1
Unit cell lengths	63.523, 122.162, 64.296
Unit cell angles	90.00, 116.46, 90.00

Refinement procedure

Resolution	41.900 - 1.950
R-factor	0.22115
Rwork	0.219
R-free	0.25746
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5kqt
RMSD bond length	0.013
RMSD bond angle	1.563
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	REFMAC (5.8.0155)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	41.900	1.990
High resolution limit [Å]	1.950	1.950
Rmerge	0.077	0.665
Number of reflections	63517
<I/σ(I)>	20.4	3.4
Completeness [%]	98.9	96.3
Redundancy	7.3	7
CC(1/2)	0.998	0.965

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		293	150 plus 150 nL drops with protein at 10 mg/mL and reservoir conditions of 20% PEG 3350, 135 mM ammonium formate. Microseeds were used for nucleation.