4PA8

Crystal structure of a de novo retro-aldolase catalyzing asymmetric Michael additions, with a covalently bound product analog

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SLS BEAMLINE X06DA
Synchrotron site	SLS
Beamline	X06DA
Temperature [K]	100
Detector technology	PIXEL
Collection date	2013-07-10
Detector	DECTRIS PILATUS 2M-F
Wavelength(s)	1.00
Spacegroup name	P 21 21 21
Unit cell lengths	62.557, 68.804, 69.341
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	46.300 - 1.200
Rwork	0.139
R-free	0.17700
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4a29
Data scaling software	Aimless (0.1.29)
Phasing software	PHASER
Refinement software	SHELXL

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	46.450	46.450	1.220
High resolution limit [Å]	1.200	6.570	1.200
Rmerge	0.031	0.026	0.811
Rpim	0.013	0.012	0.383
Total number of observations	584161	3859	23914
Number of reflections	94052
<I/σ(I)>	24.2	79.5	2.1
Completeness [%]	99.8	99.3	96.3
Redundancy	6.2	5.7	5.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5.5	277	Crystals were obtained at 4 degrees C using the sitting drop vapor diffusion method by mixing 100 nL of protein solution (13.5 mg/mL) in buffer with 100 nL of reservoir solution containing 2 M ammonium sulfate and 0.1 M BIS-TRIS buffer pH 5.5