1S20

A novel NAD binding protein revealed by the crystal structure of E. Coli 2,3-diketogulonate reductase (YiaK) NORTHEAST STRUCTURAL GENOMICS CONSORTIUM TARGET ER82

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	NSLS BEAMLINE X4A
Synchrotron site	NSLS
Beamline	X4A
Temperature [K]	100
Detector technology	CCD
Collection date	2003-06-21
Detector	ADSC QUANTUM 4
Wavelength(s)	0.9724
Spacegroup name	P 1
Unit cell lengths	75.524, 81.275, 113.215
Unit cell angles	79.55, 77.22, 82.01

Refinement procedure

Resolution	24.020 - 2.200
R-factor	0.19
Rwork	0.191
R-free	0.25000
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1NXU the apo enzyme
RMSD bond length	0.006
RMSD bond angle	1.100
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	COMO
Refinement software	CNS (1.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	24.020	2.340
High resolution limit [Å]	2.200	2.200
Rmerge	0.073	0.163
Number of reflections	125617
<I/σ(I)>	12.29	4.6
Completeness [%]	95.2	88.8
Redundancy	1.87	1.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	293	20% PEG 3350, 200 mM ammonium tartrate, and 20 mM NAD. The protein was pre-incubated with 10 mM 2,3-diketogulonate., pH 7.5, VAPOR DIFFUSION, HANGING DROP, temperature 293K