1AXE

CRYSTAL STRUCTURE OF THE ACTIVE-SITE MUTANT PHE93->TRP OF HORSE LIVER ALCOHOL DEHYDROGENASE IN COMPLEX WITH NAD AND INHIBITOR TRIFLUOROETHANOL

Experimental procedure

Source type	ROTATING ANODE
Source details	RIGAKU RUH2R
Temperature [K]	277
Detector technology	IMAGE PLATE
Collection date	1995-09-17
Detector	RIGAKU RAXIS II
Spacegroup name	P 1
Unit cell lengths	51.820, 44.580, 93.280
Unit cell angles	103.10, 87.59, 70.55

Refinement procedure

Resolution	10.000 - 2.000
R-factor	0.203
Rwork	0.203
R-free	0.26900
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2ohx
RMSD bond length	0.009
RMSD bond angle	24.100 *
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	X-PLOR (3.1)
Refinement software	X-PLOR (3.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	99.000	2.070
High resolution limit [Å]	2.000	2.000
Rmerge	0.064	0.310
Number of reflections	43756
<I/σ(I)>	5	1.3
Completeness [%]	85.2	64.6
Redundancy	1.83	1.6

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	Vapor diffusion *	8.4	4 *	CRYSTALS WERE GROWN FROM HANGING DROPS. THE INITIAL DROP CONTAINING 16MG/ML PROTEIN, A 10X EXCESS OF NAD, 5MM TRIFLUOROETHANOL AND 5% V/V OF PEG400 IN 50MM TRIS-HCL PH 8.4 WERE EQUILIBRATED AT 4 DEGREES C OVER WELLS OF SIMILAR COMPOSITION CONTAINING 15-17% PEG400., vapor diffusion - hanging drop, temperature 277K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	16 (mg/ml)
2	1	drop	NAD+	10 (M)
3	1	drop	trifluoroethanol	5 (mM)
4	1	drop	PEG400	5 (%(v/v))
5	1	reservoir	Tris-HCl	50 (mM)	pH8.4
6	1	reservoir	trifluoroethanol	5 (mM)
7	1	reservoir	PEG400	15-17 (%(v/v))