6O1C
Alpha-L-fucosidase AlfC D200A mutant in complex with 4-nitrophenyl-a-L-fucopyranoside substrate
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | SSRL BEAMLINE BL12-2 |
| Synchrotron site | SSRL |
| Beamline | BL12-2 |
| Temperature [K] | 100 |
| Detector technology | PIXEL |
| Collection date | 2019-01-14 |
| Detector | DECTRIS PILATUS3 6M |
| Wavelength(s) | 1.033170 |
| Spacegroup name | P 21 21 21 |
| Unit cell lengths | 89.734, 139.079, 263.373 |
| Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
| Resolution | 39.430 - 2.600 |
| R-factor | 0.233 |
| Rwork | 0.232 |
| R-free | 0.26070 |
| Structure solution method | MOLECULAR REPLACEMENT |
| Starting model (for MR) | 6o18 |
| RMSD bond length | 0.007 |
| RMSD bond angle | 1.385 |
| Data reduction software | XDS |
| Data scaling software | Aimless (0.7.1) |
| Phasing software | MOLREP |
| Refinement software | REFMAC (5.8.0238) |
Data quality characteristics
| Overall | Inner shell | Outer shell | |
| Low resolution limit [Å] | 39.430 | 39.430 | 2.640 |
| High resolution limit [Å] | 2.600 | 14.240 | 2.600 |
| Rmerge | 0.109 | 0.054 | 1.367 |
| Rmeas | 0.129 | 0.065 | 1.621 |
| Rpim | 0.068 | 0.035 | 0.855 |
| Total number of observations | 330144 | ||
| Number of reflections | 97990 | 615 | 4933 |
| <I/σ(I)> | 6.3 | ||
| Completeness [%] | 96.7 | 86.6 | 98.5 |
| Redundancy | 3.4 | 3.3 | 3.3 |
| CC(1/2) | 0.994 | 0.997 | 0.582 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | VAPOR DIFFUSION, SITTING DROP | 7 | 298 | 250 nL 20 mg/mL AlfC D200Aof + 250 nL mother liquor (18% w/v PEG3350, 0.1 M Bis-Tris propane, pH 7, 20 mM sodium/potassium phosphate, 1% v/v glycerol), crystals appeared after five days, were harvested in mother liquor + 20% v/v glycerol + 5 mM 4NP-Fuc for ~1 minute, flash-cooled in liquid nitrogen |
