5U5T
Crystal structure of EED in complex with H3K27Me3 peptide and 3-(benzo[d][1,3]dioxol-4-ylmethyl)piperidine-1-carboximidamide
Experimental procedure
| 実験手法 | SINGLE WAVELENGTH |
| Source type | SYNCHROTRON |
| Source details | APS BEAMLINE 17-ID |
| Synchrotron site | APS |
| Beamline | 17-ID |
| Temperature [K] | 270 |
| Detector technology | PIXEL |
| Collection date | 2013-02-22 |
| Detector | DECTRIS PILATUS 300K |
| Wavelength(s) | 0.9200 |
| Spacegroup name | P 21 21 2 |
| 格子定数 [Å] | 92.810, 177.909, 50.340 |
| 格子定数 [度] | 90.00, 90.00, 90.00 |
精密化法
| 残基 | 64.220 - 1.600 |
| R因子 | 0.17 |
| Rwork | 0.168 |
| R-free | 0.20000 |
| 結合長の平均二乗偏差(RMSD) [Å] | 0.010 |
| 結合角の平均二乗偏差(RMSD) [度] | 1.060 |
| Data reduction software | XDS |
| Data scaling software | SCALA |
| Phasing software | AMoRE |
| Refinement software | BUSTER (2.10.2) |
Quality characteristics
| Overall | |
| 分解能 [Å] (低) | 82.290 |
| 分解能 [Å] (高) | 1.600 |
| 独立反射数 | 110710 |
| <I/σ(I)> | 8 |
| 完全性 [%] | 97.2 |
| 冗長性 | 5.8 |
結晶化条件
| 結晶ID | 方法 | pH | 温度 | 溶液条件 |
| 1 | VAPOR DIFFUSION, HANGING DROP | 291 | Using a PEG/salt combination as a precipitant. Briefly, EED was incubated with 10 mM B-nicotinamide adenine dinucleotide hydrate, 2 mM of a tightly binding proprietary compound, and 0.5 mM of a synthesized peptide which comprises the helix on EZH2 which interacts with EED. The crystals were grown using the vapor diffusion method. One uL of the protein mixture was combined with 1 uL of a precipitant comprised of 20% (w/v) PEG3350, 0.2 M potassium iodide, and 0.1M Tris-HCl, pH 8.5, on a cover slip which was suspended over a reservoir comprised of 0.5 mL of precipitant at 18 Celsius and sealed. The crystals grew in 4-6 days at 18 Celsius and then harvested and soaked in defined drops consisting of 30 uL of precipitant and 2 mM of compound for 24 h. Crystals were cryopreserved for data collection using a cryosolution consisting of 30% PEG 400 (v/v), 20% (w/v) PEG 3350, 0.2 M potassium iodide, and 0.1M Tris-HCl, pH 8.5 |
