5KQT

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	2013-05-09
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.9537
Spacegroup name	P 65 2 2
Unit cell lengths	66.346, 66.346, 343.314
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	47.800 - 1.990
R-factor	0.19778
Rwork	0.196
R-free	0.23695
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3gju
RMSD bond length	0.009
RMSD bond angle	1.389
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]	47.800	2.100
High resolution limit [Å]	1.990	1.990
Rmerge	0.136	0.720
Number of reflections	32214
<I/σ(I)>	17.6	4.9
Completeness [%]	99.9	99.3
Redundancy	20.7	20.7
CC(1/2)	0.998	0.910

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	6.8	293	150 nL plus 150 nL drops of protein at 5 mg/mL and reservoir which consisted of 1.6 M ammonium sulfate, 1% dioxane, 50 mM MES pH 6.8