1H6M
Covalent glycosyl-enzyme intermediate of hen egg white lysozyme
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | ROTATING ANODE |
| Source details | RIGAKU RU200 |
| Temperature [K] | 100 |
| Detector technology | IMAGE PLATE |
| Collection date | 2000-06-15 |
| Detector | RAXIS |
| Spacegroup name | P 43 21 2 |
| Unit cell lengths | 78.632, 78.632, 36.700 |
| Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
| Resolution | 18.000 - 1.640 |
| R-factor | 0.172 |
| Rwork | 0.170 |
| R-free | 0.22200 |
| Structure solution method | MOLECULAR REPLACEMENT |
| Starting model (for MR) | REFINED COORDINATES FOR APO STRUCTURE E35Q MUTANT |
| RMSD bond length | 0.013 |
| RMSD bond angle | 1.500 * |
| Data reduction software | DENZO |
| Data scaling software | SCALEPACK |
| Phasing software | CCP4 |
| Refinement software | REFMAC (5.0) |
Data quality characteristics
| Overall | Outer shell | |
| Low resolution limit [Å] | 20.000 | 1.700 |
| High resolution limit [Å] | 1.640 | 1.640 |
| Rmerge | 0.050 | 0.190 |
| Number of reflections | 14536 | |
| <I/σ(I)> | 25 | 4.6 |
| Completeness [%] | 99.3 | 96.3 |
| Redundancy | 5.2 | 2.5 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | VAPOR DIFFUSION, HANGING DROP | 5 * | 16 * | CRYSTALS OF THE HEWL COVALENT INTERMEDIATE WERE GROWN BY THE HANGING-DROPLET VAPOUR DIFFUSION METHOD OVER 4 DAYS AT 16 C. THE DROPLET CONTAINED 2.5 UL OF A SOLUTION OF 7.5 MG/ML HEWL(E35Q), 20 MM NAG2FGLCF, 200 MM SODIUM ACETATE (NAOAC) BUFFER PH 5.0 AND 2.5 UL OF RESERVOIR SOLUTION (200MM) NAOAC, CONTAINING 4.0 % NACL (W/V), PH 4.5 WAS SUSPENDED OVER THE RESERVOIR SOLUTION. |
Crystallization Reagents in Literatures
| ID | crystal ID | solution | reagent name | concentration (unit) | details |
| 1 | 1 | drop | protein | 7.5 (mg/ml) | |
| 2 | 1 | drop | NAG2FGlcF | 20 (mM) | |
| 3 | 1 | drop | sodium acetate | 200 (mM) | |
| 4 | 1 | reservoir | sodium acetate | 200 (mM) | |
| 5 | 1 | reservoir | 4.0 (%(w/v)) |
