2BG7

Bacillus cereus metallo-beta-lactamase (BcII) Arg (121) Cys mutant. Solved at pH4.5 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	ESRF BEAMLINE ID13
Synchrotron site	ESRF
Beamline	ID13
Temperature [K]	100
Detector technology	CCD
Collection date	2001-03-30
Detector	MARRESEARCH
Spacegroup name	P 31 2 1
Unit cell lengths	67.578, 67.578, 178.769
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	6.000 - 2.100
R-factor	0.1904
Rwork	0.190
R-free	0.22200
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	1bc2
RMSD bond length	0.008
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]	59.760	2.150
High resolution limit [Å]	2.100	2.100
Rmerge	0.060	0.180
Number of reflections	28092
<I/σ(I)>	9.4	4
Completeness [%]	98.9	92.2
Redundancy	4.9	3.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	4.5	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 2.7MG/ML. DROPS WERE KEPT AT 291K AND WERE STREAK SEEDED FROM A WILD TYPE CRYSTAL AFTER 1 DAY., PH 4.50