2BG6

Bacillus cereus metallo-beta-lactamase (BcII) Arg (121) Cys mutant. Solved at pH5 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 PX14.2
Synchrotron site	SRS
Beamline	PX14.2
Temperature [K]	100
Detector technology	CCD
Collection date	2002-02-26
Detector	ADSC CCD
Spacegroup name	P 31 2 1
Unit cell lengths	67.866, 67.866, 178.470
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	6.000 - 2.300
R-factor	0.1821
Rwork	0.182
R-free	0.22270
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1bc2
RMSD bond length	0.006
RMSD bond angle	1.340
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]	41.880	2.360
High resolution limit [Å]	2.300	2.300
Rmerge	0.080	0.220
Number of reflections	21917
<I/σ(I)>	6.9	3.2
Completeness [%]	99.9	99.9
Redundancy	6.4	6.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	5	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 5.00