1E7Y

ACTIVE SITE MUTANT (D177->N) OF GLUCOSE 6-PHOSPHATE DEHYDROGENASE FROM LEUCONOSTOC MESENTEROIDES COMPLEXED WITH SUBSTRATE AND NADPH

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RUH2R
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	1998-01-15
Detector	MARRESEARCH
Spacegroup name	C 1 2 1
Unit cell lengths	129.000, 44.000, 91.100
Unit cell angles	90.00, 105.20, 90.00

Refinement procedure

Resolution	22.900 - 2.480
R-factor	0.205
Rwork	0.205
R-free	0.29600
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	REFINED COORDINATES OF APO-ENZYME IN THE SAME SPACEGROUP
RMSD bond length	0.006
RMSD bond angle	22.200 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	X-PLOR (3.851)
Refinement software	X-PLOR (3.851)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	22.900	2.590
High resolution limit [Å]	2.480	2.480
Rmerge	0.106 *	0.460 *
Total number of observations	38083 *
Number of reflections	16723
<I/σ(I)>	9	2.7
Completeness [%]	93.6	82.1
Redundancy	2.6	2.2

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5		HANGING DROP VAPOUR DIFFUSION, 2+2 MICROLITER DROPS. IN THE WELL BUFFER: 14% W/V PEG 400 IN 0.1M HEPES-NAOH, PH 7.5 WITH 0.2M CALCIUM CHLORIDE. THE PROTEIN AT 15MG/ML, WITH 2.3MM GLUCOSE 6-PHOSPHATE AND 0.6MM NADPH.

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	enzyme	15 (mg/ml)
2	1	drop	Tris-HCl	0.1 (M)
3	1	drop	NAD+	29 (mM)
4	1	reservoir	PEG400	20 (%(v/v))
5	1	reservoir	HEPES/NaOH	0.1 (M)
6	1	reservoir		0.2 (M)