4XA8

Crystal structure of D-isomer specific 2-hydroxyacid dehydrogenase from Xanthobacter autotrophicus Py2

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-G
Synchrotron site	APS
Beamline	21-ID-G
Temperature [K]	100
Detector technology	CCD
Collection date	2014-11-06
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.978
Spacegroup name	C 2 2 21
Unit cell lengths	58.762, 63.714, 154.967
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	50.000 - 1.900
R-factor	0.1655
Rwork	0.163
R-free	0.20860
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3kbo
RMSD bond length	0.013
RMSD bond angle	1.639
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0073)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.930
High resolution limit [Å]	1.900	5.160	1.900
Rmerge	0.069	0.041	0.835
Rmeas	0.073	0.043	0.881
Rpim	0.023	0.013	0.276
Total number of observations	201327
Number of reflections	22335
<I/σ(I)>	21.738		2.6
Completeness [%]	95.2	96.1	96.9
Redundancy	9	10.1	9.6
CC(1/2)		0.999	0.786

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	289	0.2 UL OF 11 MG/ML PROTEIN IN 20 MM HEPES PH 7.5, 150 MM NACL, 10% GLYCEROL, 0.1% SODIUM AZIDE, 0.5 MM TCEP and 10 mM NADH WERE MIXED WITH 0.2 UL OF THE TOP96 CONDITION #34 (0.2M MAGNESIUM CHLORIDE, 6-HYDRATE, 0.1M HEPES, 25%W/V PEG 3350 PH=7.5) AND EQUILIBRATED AGAINST 1.5 M NACL SOLUTION IN 96 WELL 3 DROP CRYSTALLIZATION PLATE (SWISSCI). BEFORE CRYSTALLIZATION PROTEIN WAS INCUBATED WITH 1/50 V/V OF 1 MG/ML TEV SOLUTION AT 289 K FOR 1 HOUR