TABLE 1.
Crystallization conditions and x-ray analysis statistics
| Data set | EtfAf | EtfAf (Hg edge)a | EtfAf + NAD+ | BcdAf |
|---|---|---|---|---|
| Crystallization | 18% PEG 3350, | 18% PEG 3350, | 17% PEG 3350, | 225% PEG1500, |
| Conditionsb | 0.1 m Bistris | 0.1 m Bistris | 0.1 m Bistris | 10% SPG buffer, |
| propane, pH 7.5 | propane, pH 7.5 | propane, pH 7.5 | pH 9.0 | |
| 0.1 m Na formate | 0.1 m Na formate | 0.1 m Na formate | ||
| 1 mm FAD | 1 mm FAD | 1 mm FAD | ||
| Protein concentration | 20 mg/ml | 20 mg/ml | 20 mg/ml | 15 mg/ml |
| Temperature | 4 °C | 4 °C | 4 °C | 4 °C |
| Soaking conditions | 0.1 mm CH3HgAc, | |||
| 5 h | ||||
| Freezing conditions | +25% | +25% | +25% | 25% |
| (w/v) glycerol | (w/v) glycerol | (w/v) glycerol | (v/v) 1,2-propandiol | |
| Data collection | ||||
| Wavelength (Å) | 1.0 | 1.006 | 1.0 | 0.9999 |
| Space group | P212121 | P212121 | P212121 | P21212 |
| Unit cell parameter | ||||
| a, b, c (Å) | 80.2, 84.9, 106.7 | 78.2, 85.7, 106.6 | 79.5, 84.8, 106.4 | 109.1, 141.4, 64.1 |
| Number of molecules per asymmetric unit | 1 | 1 | 1 | 1 |
| Resolution range (Å) | 30.0-1.6 | 30.0-3.0 | 30.0-1.45 | 30.0-1.8 |
| (highest shell) | (1.7-1.6) | (3.1-3.0) | (1.55-1.45) | (1.9-1.8) |
| Redundancy | 2.4 (2.4) | 4.6 (4.8) | 3.9 (4.0) | 7.9 (8.1) |
| Completeness (%) | 93.2 (91.8) | 99.6 (99.8) | 97.6 (96.4) | 99.9 (100) |
| Rmerge (%) | 6.9 (48.8) | 9.6 (150.9) | 4.5 (63.5) | 14.4 (78.8) |
| I/σ(I) | 11.8 (2.5) | 11.8 (1.5) | 15.6 (3.0) | 13.2 (3.9) |
| Refinement | ||||
| Resolution limit (Å) | 1.6-50.0 | 1.45-50.0 | 1.8-50.0 | |
| (highest shell) | 1.6-1.64 | 1.45-1.49 | 1.8-1.84 | |
| Rwork/Rfree (%) | 16.1/19.1 | 15.1/20.8 | 15.3/18.3 | |
| (highest shell) | 26.4/28.1 | 24.5/37.5 | 22.0/25.2 | |
| Root mean square deviation bond lengths (Å) | 0.024 | 0.023 | 0.022 | |
| Root mean square deviation bond angles (°) | 2.3 | 2.36 | 2.13 | |
| Average B (Å2) | 18.2 | 23.3 | 18.7 | |
a Phase determination. Initial SIRAS phases were obtained from native Etf data and from measurements of a crystal soaked with CH3HgAc using SHELXD (50), SHARP (51), and SOLOMON (52). After phase determination the model was built automatically at 2.5-Å resolution with PHENIX (53) supported by a model of human Etf that could be correctly placed into the unit cell and further improved at 1.6-Å resolution with ARP/WARP (54).
b Applied crystallization screens: JBScreen HTS I+II, pentaerythritol, PACT++ (JBS), JCSG Core Suite I–IV (Qiagen), and PGA (MDL).