2BGA

Bacillus cereus metallo-beta-lactamase (BcII) Arg (121) Cys mutant. Solved at pH7 using 20 Micromolar ZnSO4 in the buffer. 1mM DTT was used as a reducing agent. Cys221 is oxidized.

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	SRS BEAMLINE PX9.6
Synchrotron site	SRS
Beamline	PX9.6
Temperature [K]	100
Detector technology	CCD
Collection date	2002-04-19
Detector	ADSC CCD
Spacegroup name	P 31 2 1
Unit cell lengths	67.391, 67.391, 178.530
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	6.000 - 2.700
R-factor	0.1856
Rwork	0.186
R-free	0.24750
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1bc2
RMSD bond length	0.007
RMSD bond angle	1.400
Data reduction software	DENZO
Data scaling software	SCALA
Phasing software	CNS
Refinement software	CNS (1.1)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	2.770	2.770
High resolution limit [Å]	2.700	2.700
Rmerge	0.160	0.560
Number of reflections	13382
<I/σ(I)>	4	1.2
Completeness [%]	99.4	99.4
Redundancy	2.9	2.9

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7	291	PROTEIN WAS CRYSTALLISED USING HANGING DROP VAPOR DIFFUSION. RESERVOIR SOLUTION CONTAINED 100MM TRIS AT PH4.5-5, 70-75% AMMONIUM SULPHATE, 1MM DTT OR 1MM DTT AND 1MM TCEP-HCL, 2MM ZNSO4 AND 0.1% AZIDE. PROTEIN CONCENTRATION OF 2.7 MG/ML. DROPS WERE KEPT AT 291K AND WERE STREAK SEEDED FROM A WILD TYPE CRYSTAL AFTER 1 DAY., pH 7.00