5KR4

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 21 21 21
Unit cell lengths	67.053, 121.783, 125.629
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	45.800 - 2.000
R-factor	0.19381
Rwork	0.192
R-free	0.22415
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5kqt
RMSD bond length	0.013
RMSD bond angle	1.565
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]	45.800	2.040
High resolution limit [Å]	2.000	2.000
Rmerge	0.156	0.726
Number of reflections	70329
<I/σ(I)>	10.4	2.4
Completeness [%]	99.8	97.1
Redundancy	7.3	7
CC(1/2)	0.993	0.756

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 12% PEG 8000, 15% glycerol, 150 mM MgCl2, 100 mM Tris pH 7.1. Microseeds were used for nucleation of the crystals.