4WEQ

Crystal structure of NADPH-dependent glyoxylate/hydroxypyruvate reductase SMc02828 (SmGhrA) from Sinorhizobium meliloti in complex with NADP and sulfate

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 32 2 1
Unit cell lengths	108.153, 108.153, 80.657
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	50.000 - 2.000
R-factor	0.1561
Rwork	0.154
R-free	0.19620
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.015
RMSD bond angle	1.754
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	MOLREP
Refinement software	REFMAC (5.8.0073)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	2.030
High resolution limit [Å]	2.000	5.430	2.000
Rmerge	0.069	0.042	0.846
Rmeas	0.072	0.045	0.883
Rpim	0.021	0.013	0.252
Total number of observations	452447
Number of reflections	37017
<I/σ(I)>	9.4
Completeness [%]	100.0	99.6	100
Redundancy	12.2	11.5	12.2
CC(1/2)		0.998	0.920

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		289	0.2 ul of 12 mg/ml protein in 20 mM HEPES pH 7.5, 150 mM NaCl, 10% Glycerol, 0.1% Sodium Azide, 0.5 mM TCEP, and 5 mM NADP were mixed with 0.2 ul of the MCSG-I condition #6 (0.2 M Ammonium Sulfate 0.1 M Bis-Tris:HCl pH 5.5 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-ligand mixture was incubated with 1/40 v/v of 2 mg/ml chymotrypsin solution at 289 K for 3 hours