1DD6

IMP-1 METALLO BETA-LACTAMASE FROM PSEUDOMONAS AERUGINOSA IN COMPLEX WITH A MERCAPTOCARBOXYLATE INHIBITOR

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU300
Temperature [K]	298
Detector technology	IMAGE PLATE
Collection date	1997-04-21
Detector	MARRESEARCH
Spacegroup name	P 21 21 21
Unit cell lengths	49.300, 51.200, 203.300
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	20.000 - 2.000
R-factor	0.198
Rwork	0.198
R-free	0.25900
RMSD bond length	0.018
RMSD bond angle	2.000
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	AMoRE
Refinement software	CNS

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	20.000	2.070
High resolution limit [Å]	2.000	2.000
Rmerge	0.103	0.314
Total number of observations	545270 *
Number of reflections	34187 *
<I/σ(I)>	21
Completeness [%]	95.9 *	97.6
Redundancy	16	7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5 *	293	SITTING DROPS WERE PREPARED BY MIXING EQUAL VOLUMES OF PROTEIN WHICH WAS 14 MG/ ML IN 20MM HEPES, PH 7.5 AND TO WHICH AN EXCESS OF SOLID INHIBITOR HAD BEEN ADDED AND RESERVOIR SOLUTION (30% PEG 2000 MONOMETHYLETHER, 0.1M SODIUM ACETATE, PH 5.0 AND 0.2M AMMONIUM SULFATE). THIS MIXTURE WAS INCUBATED OVERNIGHT AT 4C AND CENTRIFUGED TO REMOVE PRECIPITATE BEFORE SETTING UP CRYSTALLIZATION DROPS. CO-CRYSTALS WERE GROWN FROM 6 ML SITTING DROPS OF THE PROTEIN-RESERVOIR SOLUTION AND 0.3 ML OF THE RESERVOIR SOLUTION AT EITHER ROOM TEMPERATURE OR 4C, pH 6.5, VAPOR DIFFUSION, SITTING DROP, temperature 293K
1	VAPOR DIFFUSION, SITTING DROP	7.5 *	293	SITTING DROPS WERE PREPARED BY MIXING EQUAL VOLUMES OF PROTEIN WHICH WAS 14 MG/ ML IN 20MM HEPES, PH 7.5 AND TO WHICH AN EXCESS OF SOLID INHIBITOR HAD BEEN ADDED AND RESERVOIR SOLUTION (30% PEG 2000 MONOMETHYLETHER, 0.1M SODIUM ACETATE, PH 5.0 AND 0.2M AMMONIUM SULFATE). THIS MIXTURE WAS INCUBATED OVERNIGHT AT 4C AND CENTRIFUGED TO REMOVE PRECIPITATE BEFORE SETTING UP CRYSTALLIZATION DROPS. CO-CRYSTALS WERE GROWN FROM 6 ML SITTING DROPS OF THE PROTEIN-RESERVOIR SOLUTION AND 0.3 ML OF THE RESERVOIR SOLUTION AT EITHER ROOM TEMPERATURE OR 4C, pH 6.5, VAPOR DIFFUSION, SITTING DROP, temperature 293K

Crystallization Reagents in Literatures

ID	crystal ID	solution	reagent name	concentration (unit)	details
1	1	drop	protein	14 (mg/ml)
2	1	drop	HEPES	20 (mM)	pH7.5
3	1	reservoir	PEG MME2000	30 (%)
4	1	reservoir	sodium acetate	0.1 (M)	pH5.6
5	1	reservoir	ammonium sulfate	0.2 (M)