Table 1.
Crystallization conditions and data collection statistics.
| Structure | TTR-Fe | TTR-Mn | TTR-Cu | TTR-Cu-Aβ |
|---|---|---|---|---|
| PDB code | 5N5Q | 5N62 | 5N7C | Not deposited |
| Metal-Ligand | Fe | Mn | Cu | Cu-Aβ |
| Crystallization | 27% polyethylene glycol 4,000 (PEG4K), 0.2 M imidazole malate, pH 6.0 | 26% PEG4K, 0.2 M imidazole malate, pH 6.0 | 21% PEG4K, 0.14 M imidazole malate, pH 6.0, 3.6% MPEG 5 K, 0.03 M sodium acetate, pH 5.5 | Co-crystallization with Aβ 1–28 and CuCl2 in 21% PEG4K, 0.14 M imidazole malate, pH 6.0, 3.6% MPEG, 5 K, 0.1 M sodium acetate, pH 5.5 |
| Cryoprotectant | 40% SM3*, 25% MPEG 5 K, 0.1 M CHC (90% acid/10% basic), 30 mM FeCl2, 2 h soak. | 40% CM7*, 25% MPEG 5 K, 5 mM MnCl2, 10 min soak. | 40% SM3*, 25% MPEG 5 K, 0.1 M CHC(90% acid/10% basic), 30 mM CuCl2 30 min soak. | 40% CM1*, 25% MPEG 5 K, 0.1 M CHC (90% acid/10% basic), 30 mM CuCl2 30 min soak. |
| Data Collection | anomalous | anomalous | anomalous | non anomalous |
| Source | Soleil Proxima-2 | Soleil Proxima-2 | Soleil Proxima-2 | Soleil Proxima-2 |
| Wavelength (Å) | 1.739153 | 1.891993 | 1.175919 | 0.980035 |
| Space group | P21212 | P21212 | P21212 | P21212 |
| Unit-cell (Å) | 42.82 83.10 65.34 | 43.27 85.83 63.90 | 42.99 82.54 67.77 | 43.10 82.75 67.14 |
| Molec./asym. | 2 | 2 | 2 | 2 |
| Resolution (Å) | 50–2.53/2.68–2.53 | 50–1.8/1.85–1.80 | 50–2.45/2.59–2.45 | 50–2.14/2.27–2.14 |
| CC1/2 (%) | 99.6/30.0 | 99.8/39.0 | 99.8/89.0 | 99.9/52.5 |
| 〈I/σ(I)〉 | 8.28/0.83 | 11.53/1.52 | 14.75/3.29 | 13.0/1.10 |
| R-merge (%) | 20.0/198 | 17.6/217 | 10.7/67.2 | 12.9/215 |
| R-factor (%) | 18.5/182 | 16.3/202 | 9.9/62.1 | 12.4/222 |
| Completeness (%) | 99.2/95.3 | 100/100 | 99.5/96.7 | 99.8/98.9 |
| Multiplicity | 13.21/6.87 (anomalous) | 7.37 (anomalous) | 13.7/7.0 (anomalous) | 13.1 |
| Refinement | REFMAC5 | Phenix | REFMAC5 | REFMAC5 |
| Resolution (Å) | 40.91–2.53/2.60–2.53 | 38.63–1.80/1.89–1.80 | 41.06–2.45/2.51–2.45 | 38.23–2.43/2.49–2.43 |
| No. of reflections | 8113/548 (non-anomalous) | 22723/2658 (anomalous) | 8868/467 (non-anomalous) | 9042/650 (non-anomalous) |
| R-work | 19.2/41.0 | 19.5/28.4 | 16.9/20.3 | 20.9/31.0 |
| R-free | 25.3/40.4 | 24.4/33.2 | 25.4/43.1 | 29.7/44.4 |
| RMSD Bond lengths (Å) | 0.013 | 0.009 | 0.009 | 0.015 |
| RMSD Bond angles (°) | 1.6 | 1.025 | 1.714 | 1.773 |
| Ramachandran favored | 96.0% | 98.0% | 94.0% | 96.9% |
| Ramachandran outliers | 0 | 0 | 0 | 3 |
Cryoprotection: SM1: 12.5% diethylene glycol + 12.5% glycerol + 12.5% 1,2-propanediol + 25% DMSO + 25% 1,4-dioxane; SM3: 25% diethylene glycol + 25% ethylene glycol + 25% glycerol + 25% 1,4-dioxane; CM1: 12.5% diethylene glycol + 37.5% 1,2-propanediol + 12.5% DMSO; CM7: 12.5% di-ethylene glycol + 12.5% ethylene glycol + 12.5% glycerol + 25% 1,2-propanediol + 12.5% DMSO68. Cryoprotectant solution is formulated with 40% v/v mixed compounds (CM), 50% v/v precipitant and 10% v/v buffer69. CHC: Linear mixed buffer composed of citric acid, HEPES and CHES; acid mix at pH 4.0, basic at pH 10.070. CC1/2: Data quality correlation coefficient71. Data collection statistics are from XDS60. Refinement statistics are from REFMAC563 or phenix.refine (Phenix)65.