1W4N

AGAO covalent complex with Tranylcypromine

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU200
Temperature [K]	100
Detector technology	CCD
Collection date	2003-06-05
Detector	MARRESEARCH
Spacegroup name	I 1 2 1
Unit cell lengths	158.113, 63.001, 184.490
Unit cell angles	90.00, 112.04, 90.00

Refinement procedure

Resolution	28.280 - 1.650
R-factor	0.172
Rwork	0.171
R-free	0.19900
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1av4
RMSD bond length	0.011
RMSD bond angle	1.405
Data reduction software	DENZO
Data scaling software	SCALEPACK
Refinement software	REFMAC (5.1.24)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	28.250	1.670
High resolution limit [Å]	1.650	1.650
Rmerge	0.060	0.460
Number of reflections	200663
<I/σ(I)>	15.73	3
Completeness [%]	98.2	80
Redundancy	7.4

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 SINGLE CRYSTAL WAS SOAKED WITH TCP BY SLOWLY ADDING TCP TO THE DROP TO ACHIEVE A 100 FOLD EXCESS OF TCP OVER ENZYME ACTIVE SITES. THE CRYSTAL WAS CRYOPROTECTED BY GRADUAL INCREMENTAL SOAKING IN RESERVOIR SOLUTION MIXED WITH GLYCEROL, WITH THE CONCENTRAION OF TCP BEING MAINTAINED CONSTANT. THE CONCENTRATION OF GLYCEROL WAS INCREASED FROM 0 TO 30% IN 2-3% INCREMENTS DURING 2 HOURS.