5J23

Crystal structure of NADPH-dependent glyoxylate/hydroxypyruvate reductase SMc04462 (SmGhrB) from Sinorhizobium meliloti in complex with 2'-phospho-ADP-ribose

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)	0.97856
Spacegroup name	H 3
Unit cell lengths	175.822, 175.822, 136.839
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	36.670 - 2.300
R-factor	0.1475
Rwork	0.146
R-free	0.16900
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4z0p
RMSD bond length	0.013
RMSD bond angle	1.491
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	HKL-3000
Refinement software	REFMAC (5.8.0135)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	2.340
High resolution limit [Å]	2.300	6.240	2.300
Rmerge	0.077	0.030	0.662
Number of reflections	70083
<I/σ(I)>	7.5		1.8
Completeness [%]	100.0	99.7	100
Redundancy	3.9	3.8	3.9

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7	289	0.2 ul of 13 mg/ml protein in 10 mM NADPH, 10 mM Glycolic Acid, 20 mM HEPES pH 7.5, 150 mM NaCl, 10% Glycerol, 0.1% Sodium Azide and 0.5 mM TCEP were mixed with 0.2 ul of the MCSG Suite 2 condition #20 (1.1 M Malonic acid, 0.072 M Succinic acid, 0.15M Ammonium dihydrogen citrate, 0.18 M DL-Malic Acid, 0.096 M Ammonium tartrate, 0.24 M Sodium acetate anhydrous, 0.3 M Sodium formate, pH=7.0) and equilibrated against 1.5 M NaCl solution in 96 Well 3 drop Crystallization Plate (Swissci). Before crystallization, the protein was incubated with 1/15 v/v of 1 mg/ml rTEV solution at 289 K for 3 hours.