5TX7

Crystal structure of D-isomer specific 2-hydroxyacid dehydrogenase from Desulfovibrio vulgaris

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	C 2 2 21
Unit cell lengths	91.109, 117.107, 135.840
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	50.000 - 2.510
R-factor	0.1768
Rwork	0.174
R-free	0.22980
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2cuk
RMSD bond length	0.009
RMSD bond angle	1.362
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	BALBES
Refinement software	REFMAC (5.8.0151)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.010	50.000	2.540
High resolution limit [Å]	2.500	6.780	2.500
Rmerge	0.091	0.058	0.770
Number of reflections	25228
<I/σ(I)>	8.5		2.53
Completeness [%]	99.9	99	100
Redundancy	5.2	4.9	5.2
CC(1/2)		0.994	0.666

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5.5	289	0.2 ul of 19 mg/ml protein in 50 mM Tris pH 7.5, 300 mM NaCl, and 0.5 mM TCEP were mixed with 0.2 ul of the MCSG II condition #21 (1M Ammonium Sulfate, 0.1M Bis-Tris, 1%w/v PEG 3350 pH=5.5) and equilibrated against 1.5 M NaCl solution in 96 Well 3 drop Crystallization Plate (Swissci). Before crystallization protein was incubated with 1/15 v/v of 1 mg/ml rTEV solution at 289 K for 3 hours