4XCV

Probable 2-hydroxyacid dehydrogenase from Rhizobium etli CFN 42 in complex with NADPH

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 19-ID
Synchrotron site	APS
Beamline	19-ID
Temperature [K]	100
Detector technology	CCD
Collection date	2014-07-23
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.97912
Spacegroup name	P 41 21 2
Unit cell lengths	65.663, 65.663, 151.458
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	50.000 - 1.400
R-factor	0.1507
Rwork	0.149
R-free	0.17750
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3kbo
RMSD bond length	0.014
RMSD bond angle	1.748
Data scaling software	HKL-3000
Phasing software	HKL-3000
Refinement software	REFMAC (5.8.0073)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.420
High resolution limit [Å]	1.400	3.800	1.400
Rmerge	0.067	0.038	0.914
Rmeas	0.072	0.040	0.986
Rpim	0.025	0.014	0.361
Total number of observations	426874
Number of reflections	62557
<I/σ(I)>	10.3		2.1
Completeness [%]	94.8	91.4	89
Redundancy	6.8	8.6	6.4
CC(1/2)		0.998	0.777

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	289	0.2 ul of 15 mg/ml protein in 20 mM HEPES pH 7.5, 150 mM NaCl, 10% Glycerol, 0.1% Sodium Azide, 0.5 mM TCEP 10 mM NADP and 10 mM glycolic acid were mixed with 0.2 ul of the MCSG-I condition #45 (0.2 M Sodium Chloride, 0.1 M Tris:HCl pH 8.5, 25% (w/v) PEG 3350) and equilibrated against 1.5 M NaCl solution in 96 Well 3 drop Crystallization Plate (Swissci). Before crystallization the protein was incubated with 1/50 v/v of 2 mg/ml TEV solution at 289 K.