1E7M

ACTIVE SITE MUTANT (D177->N) OF GLUCOSE 6-PHOSPHATE DEHYDROGENASE FROM LEUCONOSTOC MESENTEROIDES

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RUH2R
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	1997-12-15
Detector	MARRESEARCH
Spacegroup name	C 1 2 1
Unit cell lengths	130.300, 44.400, 92.800
Unit cell angles	90.00, 106.70, 90.00

Refinement procedure

Resolution	24.200 - 2.540
R-factor	0.222
Rwork	0.222
R-free	0.29900
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	22.300 *
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]	30.000	2.590
High resolution limit [Å]	2.540	2.540
Rmerge	0.086 *	0.259 *
Total number of observations	46259 *
Number of reflections	15753
<I/σ(I)>	12.7	3.3
Completeness [%]	95.1	85.6
Redundancy	3.2

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: 20% W/V PEG 400 IN 0.1M HEPES-NAOH, PH 7.5 WITH 0.2M CALCIUM CHLORIDE. THE PROTEIN AT 15MG/ML, WITH 29MM NAD+.

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)