1REQ
METHYLMALONYL-COA MUTASE
Experimental procedure
| Source type | SYNCHROTRON |
| Source details | ESRF BEAMLINE ID2 |
| Synchrotron site | ESRF |
| Beamline | ID2 |
| Detector technology | IMAGE PLATE |
| Collection date | 1994-06-24 |
| Detector | MAR scanner 300 mm plate |
| Spacegroup name | P 1 21 1 |
| Unit cell lengths | 119.800, 161.300, 88.400 |
| Unit cell angles | 90.00, 105.10, 90.00 |
Refinement procedure
| Resolution | 20.000 - 2.000 |
| R-factor | 0.22 * |
| Rwork | 0.220 |
| R-free | 0.27500 |
| RMSD bond length | 0.017 |
| RMSD bond angle | 0.039 |
| Data reduction software | MOSFLM |
| Refinement software | REFMAC |
Data quality characteristics
| Overall | |
| Low resolution limit [Å] | 20.000 |
| High resolution limit [Å] | 2.000 |
| Rmerge | 0.051 |
| Total number of observations | 1008268 * |
| Number of reflections | 217377 |
| Completeness [%] | 99.8 |
| Redundancy | 4.6 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | Vapor diffusion * | 7.5 * | 23 * | THE CRYSTALS WERE GROWN IN THE PRESENCE OF 2MM EXCESS 5'-DEOXYADENOSYLCOBALAMIN (COENZYME B12) AND 12MM DESULPHO-COA (FINAL CONCENTRATIONS), EQUILIBRATED AGAINST 14% W/V PEG 4000 AND 20% V/V GLYCEROL, 100MM TRIS PH 7.5. THE ELECTRON DENSITY MAPS SHOW NO ADENOSYL GROUP ATTACHED TO THE COBALT ATOM, AND SPECTRA FROM SIMILAR CRYSTALS SHOW EVIDENCE OF SUBSTANTIAL REDUCTION OF COIII TO COII, WHICH IS 5-COORDINATE. THE COBALAMIN IN THIS CRYSTAL STRUCTURE IS BEST CONSIDERED AS REDUCED COB(II)ALAMIN (OR B12R). THE BOND LENGTH FROM THE COBALT TO THE LOWER AXIAL LIGAND, NE2 OF HIS A 610 (OR HIS C 610) IS SIGNIFICANTLY LONGER THAN THAT IN MODEL COMPOUNDS. POORLY ORDERED LOOPS: DENSITY IN THE FOLLOWING REGIONS IS POOR, AND THE MODEL MUST BE CONSIDERED UNRELIABLE. ALPHA CHAIN: A 1, C 1 MISSING, A 2 - A 3, C 2 - C 3 WEAK. BETA CHAIN: B 1 - C 19, C 1 - C 16 MISSING; B 184 - B 191, D 184 - D 191 WEAK; D 228 - D 230 WEAK; D 271 - D 276 VERY POOR DENSITY (MUCH BETTER IN CHAIN B); D 315 WEAK; D 474 - D 428 WEAK. |
Crystallization Reagents in Literatures
| ID | crystal ID | solution | reagent name | concentration (unit) | details |
| 1 | 1 | reservoir | PEG4000 | 14 (%(w/v)) | |
| 2 | 1 | reservoir | glycerol | 20 (%(v/v)) | |
| 3 | 1 | reservoir | Tris-HCl | 100 (mM) | |
| 4 | 1 | drop | PEG4000 | 14 (%(w/v)) | |
| 5 | 1 | drop | glycerol | 20 (%(v/v)) | |
| 6 | 1 | drop | Tris-HCl | 100 (mM) | |
| 7 | 1 | drop | protein | 20 (mg/ml) | |
| 8 | 1 | drop | coenzyme B12 | 2 (mM) | |
| 9 | 1 | drop | desulpho-CoA | 12 (mM) |
