5V7N

Crystal structure of NADPH-dependent glyoxylate/hydroxypyruvate reductase SMc04462 (SmGhrB) from Sinorhizobium meliloti in complex with NADP and 2-Keto-D-gluconic acid

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-F
Synchrotron site	APS
Beamline	21-ID-F
Temperature [K]	100
Detector technology	CCD
Collection date	2016-11-11
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	0.97872
Spacegroup name	H 3
Unit cell lengths	175.902, 175.902, 135.336
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	45.110 - 1.750
R-factor	0.1451
Rwork	0.144
R-free	0.16660
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5j23
RMSD bond length	0.011
RMSD bond angle	1.502
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0158)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	1.780
High resolution limit [Å]	1.750	4.750	1.750
Rmerge	0.113	0.073	0.878
Rmeas	0.129	0.084	0.987
Rpim	0.060	0.041	0.447
Number of reflections	156897		7876
<I/σ(I)>	8.6		1.8
Completeness [%]	99.6	97.3	100
Redundancy	4.9	4.3	4.9
CC(1/2)		0.990	0.607

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5.9	289	0.2 uL 13 mg/mL protein in 20 mM HEPES, pH 7.5, 150 mM sodium chloride, 10% glycerol, 0.1% sodium azide, 0.5 mM TCEP, 5 mM NADP, 100 mM 2-keto-D-gluconic acid hemicalcium salt + 0.2 uL TOP96 condition #24 (0.2 M magnesium formate, pH 5.9, 20% w/v PEG3350), equilibrated against 1.5 M sodium chloride in a 96-well, 3-drop crystallization plate (Swissci), incubated with 1/20 v/v 1 mg/mL rTEV at 289 K for 3 hours prior to crystallization