5HI7
Co-crystal structure of human SMYD3 with an aza-SAH compound
Experimental procedure
| Experimental method | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | APS BEAMLINE 21-ID-D |
| Synchrotron site | APS |
| Beamline | 21-ID-D |
| Temperature [K] | 295 |
| Detector technology | CCD |
| Collection date | 2013-01-13 |
| Detector | MARMOSAIC 300 mm CCD |
| Wavelength(s) | 1.07820 |
| Spacegroup name | P 21 21 21 |
| Unit cell lengths | 60.470, 66.401, 104.851 |
| Unit cell angles | 90.00, 90.00, 90.00 |
Refinement procedure
| Resolution | 44.709 - 2.150 |
| R-factor | 0.1999 |
| Rwork | 0.198 |
| R-free | 0.22570 |
| Structure solution method | MOLECULAR REPLACEMENT |
| RMSD bond length | 0.003 |
| RMSD bond angle | 0.626 |
| Data reduction software | HKL-2000 |
| Data scaling software | HKL-2000 |
| Phasing software | PHENIX |
| Refinement software | PHENIX (dev_1801) |
Data quality characteristics
| Overall | |
| Low resolution limit [Å] | 50.000 |
| High resolution limit [Å] | 2.100 |
| Number of reflections | 46288 |
| <I/σ(I)> | 11.4 |
| Completeness [%] | 99.2 |
| Redundancy | 2.2 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | VAPOR DIFFUSION, SITTING DROP | 295 | SMYD3 crystallization and soaking of GSK2807: SMYD3 (1-428) expressed in baculovirus and with SAH in the SAM-binding site was crystallized in sitting drops at 22C using 1uL fresh protein at 8.9mg/mL with the addition of 1uL mother liquor (200mM MgOAc Tetrahydrate, 20% PEG 3350). Crystals grew to a large size but varied in quantity when set up in replicates. A seed stock was formed from these crystals and added at 20% to the mother liquor solution prior to plate setup. Crystals of large size were then created with 1.3uL protein and 1.3uL of mother liquor containing seeds in a sitting drop MRC-2 plate (Hampton Research). Crystal nucleation occurred overnight and reached maximum size in 4 days |






