2E7F
5-methyltetrahydrofolate corrinoid/iron sulfur protein methyltransferase complexed with methyltetrahydrofolate to 2.2 Angsrom resolution
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | NSLS BEAMLINE X12C |
Synchrotron site | NSLS |
Beamline | X12C |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 2000-07-30 |
Detector | BRANDEIS - B4 |
Wavelength(s) | 0.91938 |
Spacegroup name | P 21 21 21 |
Unit cell lengths | 50.159, 78.547, 135.617 |
Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
Resolution | 30.880 - 2.200 |
R-factor | 0.178 |
Rwork | 0.172 |
R-free | 0.23000 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 1f6y |
RMSD bond length | 0.006 |
RMSD bond angle | 1.018 |
Data reduction software | MOSFLM |
Data scaling software | CCP4 ((SCALA)) |
Phasing software | EPMR |
Refinement software | CNS (1.1) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 30.920 | 2.260 |
High resolution limit [Å] | 2.200 | 2.200 |
Rmerge | 0.086 | |
Number of reflections | 26806 | |
<I/σ(I)> | 11.5 | 3.5 |
Completeness [%] | 96.2 | 87.9 |
Redundancy | 3.8 | 3.5 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, HANGING DROP | 7.5 | 298 | 8-15% PEGmme 5000, 0.05M Calcium acetate, 20% Glycerol, 0.05M HEPES buffer; 3-fold molar excess of the MTHF substrate (Schricks Labolatories, Jona, Switzerland). The supersaturation of the precipitant solution required dilution of the protein-CH3-H4folate complex by 50-100-fold in order to obtain single crystals., pH 7.5, VAPOR DIFFUSION, HANGING DROP, temperature 298K |