2BG2
Bacillus cereus metallo-beta-lactamase (BcII) Arg (121) Cys mutant. Solved at pH4.5 using 20mM ZnSO4 in the buffer. 1mM DTT and 1mM TCEP- HCl were used as reducing agents. Cys221 is reduced.
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 | 2002-05-31 |
Detector | ADSC CCD |
Spacegroup name | P 31 2 1 |
Unit cell lengths | 67.300, 67.300, 177.183 |
Unit cell angles | 90.00, 90.00, 120.00 |
Refinement procedure
Resolution | 6.000 - 2.400 |
R-factor | 0.1787 |
Rwork | 0.179 |
R-free | 0.22440 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 1bc2 |
RMSD bond length | 0.007 |
RMSD bond angle | 1.320 |
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] | 58.720 | 2.460 |
High resolution limit [Å] | 2.400 | 2.400 |
Rmerge | 0.090 | 0.370 |
Number of reflections | 18697 | |
<I/σ(I)> | 5.7 | 1.8 |
Completeness [%] | 99.0 | 99 |
Redundancy | 5.2 | 5.2 |
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 2.7 MG/ML. DROPS WERE KEPT AT 291K AND WERE STREAK SEEDED FROM A WILD TYPE CRYSTAL AFTER 1 DAY., pH 4.50 |