5BQF

Probable 2-hydroxyacid dehydrogenase from Rhizobium etli CFN 42 in complex with NADP, HEPES and L(+)-tartaric acid

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 19-ID
Synchrotron site	APS
Beamline	19-ID
Temperature [K]	300
Detector technology	CCD
Collection date	2014-11-08
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.979
Spacegroup name	P 41 21 2
Unit cell lengths	65.692, 65.692, 151.406
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	50.000 - 1.450
R-factor	0.1398
Rwork	0.139
R-free	0.16540
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4xcv
RMSD bond length	0.008
RMSD bond angle	1.441
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	HKL-3000
Refinement software	REFMAC (5.8.0103)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.480
High resolution limit [Å]	1.450	3.940	1.450
Rmerge	0.063	0.039	0.547
Rmeas	0.072	0.044	0.633
Rpim	0.035	0.021	0.313
Total number of observations	231378
Number of reflections	58208
<I/σ(I)>	10.9		1.9
Completeness [%]	97.5	88.6	99.5
Redundancy	4	3.9	3.9
CC(1/2)		0.998	0.759

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	300	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 were mixed with 0.2 ul of the MCSG Suite 2 condition #71 (0.2M Na tartrate, 20%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