1W5Z

AGAO covalent complex with Benzylhydrazine

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU200
Temperature [K]	100
Detector technology	CCD
Collection date	2003-03-25
Detector	MARRESEARCH
Spacegroup name	C 1 2 1
Unit cell lengths	158.126, 62.665, 92.283
Unit cell angles	90.00, 112.07, 90.00

Refinement procedure

Resolution	29.850 - 1.860
R-factor	0.154
Rwork	0.152
R-free	0.18700
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1av4
RMSD bond length	0.013
RMSD bond angle	1.465
Data reduction software	DENZO
Data scaling software	SCALEPACK
Refinement software	REFMAC (5.1.24)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	29.860	1.930
High resolution limit [Å]	1.860	1.860
Rmerge	0.060	0.410
Number of reflections	65983
<I/σ(I)>	9.46	2.13
Completeness [%]	94.2	83
Redundancy	3.033	2.69

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7	293	AGAO CRYSTALS WERE GROWN IN ABOUT 2 WEEKS BY HANGING-DROP VAPOR DIFFUSION AT 293K. THE RESERVOIR WAS 1.0-1.5M (NH4)2SO4, 2MM CUSO4, 0.15M NA CITRATE, PH 6.0-7.5. THE DROPS WERE 1.5MICROL OF 11.7 MG/ML PROTEIN, 0.05M HEPES, PH 7.0, PLUS 1.5MICROL OF RESERVOIR SOLUTION. A CRYSTAL WAS CRYOPROTECTED BY GRADUAL INCREMENTAL SOAKING IN RESERVOIR SOLUTION MIXED WITH GLYCEROL. THE CONCENTRATION OF GLYCEROL WAS INCREASED FROM 0 TO 30% IN 2-3% INCREMENTS DURING 2 HOURS. FOLLOWING CRYOPROTECTION, THE CRYSTAL WAS SOAKED FOR 15 MINUTES IN CRYOPROTECTANT WITH THE ADDITION OF 2 MILLIMOLAR BENZYLHYDRAZINE DIHYDROCHLORIDE (SIGMA). THE CRYSTAL WAS THEN FROZEN IN THE CRYOSTREAM.