3DUF
Snapshots of catalysis in the E1 subunit of the pyruvate dehydrogenase multi-enzyme complex
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | ESRF BEAMLINE ID23-1 |
| Synchrotron site | ESRF |
| Beamline | ID23-1 |
| Temperature [K] | 100 |
| Detector technology | CCD |
| Collection date | 2005-11-11 |
| Detector | MARMOSAIC 225 mm CCD |
| Wavelength(s) | 0.9795 |
| Spacegroup name | P 1 21 1 |
| Unit cell lengths | 69.247, 231.995, 92.614 |
| Unit cell angles | 90.00, 90.74, 90.00 |
Refinement procedure
| Resolution | 59.440 - 2.500 |
| R-factor | 0.19233 |
| Rwork | 0.188 |
| R-free | 0.26303 |
| Structure solution method | MOLECULAR REPLACEMENT |
| Starting model (for MR) | 1w85 |
| RMSD bond length | 0.008 |
| RMSD bond angle | 1.282 |
| Data reduction software | MOSFLM |
| Data scaling software | SCALA |
| Phasing software | AMoRE |
| Refinement software | REFMAC (5.2.0019) |
Data quality characteristics
| Overall | Outer shell | |
| Low resolution limit [Å] | 66.000 | 2.640 |
| High resolution limit [Å] | 2.500 | 2.500 |
| Rmerge | 0.111 | 0.260 |
| Number of reflections | 99351 | |
| <I/σ(I)> | 3.1 | 5.4 |
| Completeness [%] | 98.9 | 99.9 |
| Redundancy | 3.7 | 3.7 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | VAPOR DIFFUSION, SITTING DROP | 5.5 | 291.15 | The protein solution was then mixed in 1:1 volume ratio of crystallization buffer consisting of 8-12 % mono-methyl ether polyethylene glycol (MME PEG) 5000, 0.1 M Na maleate pH 5.5, and the droplet was left to equilibrate against a reservoir of neat crystallization buffer., VAPOR DIFFUSION, SITTING DROP, temperature 291.15K |
