Table 1.
Crystallization conditions, data collection, and structure refinement
| AlgM1M2SS/AlgQ2 + 6–8M | AlgQ2 + 7–10M | AlgM1M2SS/AlgQ2 + Δ3M | AlgQ2 + Δ3M | |
|---|---|---|---|---|
| Crystallization conditions | ||||
| Sample solution | 7 mg/ml AlgM1(d24) M2(H10)SS(E160Q) | 7 mg/ml AlgM1(d24) M2(H10)SS(E160Q) | ||
| 3.5 mg/ml AlgQ2 | 3 mg/ml AlgQ2 | 3.5 mg/ml AlgQ2 | 3 mg/ml AlgQ2 | |
| 2 mm 6–8M | 1 mm 7–10M | 2 mm Δ3M | 1 mm Δ3M | |
| 8 mm CHAPSO | 16 mm CHAPSO | |||
| Reservoir solution | 22% PEG 3000 | 25% PEG 4000 | 22% PEG 3000 | 30% PEG 4000 |
| 0.1 m ADA-NaOH (pH 6.6) | 0.1 m Tris-HCl (pH 8.5) | 0.1 m ADA-NaOH (pH 6.6) | 0.1 m Tris-HCl (pH 8.5) | |
| 0.15 m HCOONa | 0.2 m CaCl2 | 0.15 m NaCl | 0.2 m Li2SO4 | |
| Data collection | ||||
| Beam line | SPring-8 BL38B1 | SPring-8 BL26B2 | SPring-8 BL38B1 | SPring-8 BL38B1 |
| Detector | MX225HE (Rayonix) | MX225 (Rayonix) | Quantum315r (Area Detector Systems Corp.) | MX225HE (Rayonix) |
| Detector–crystal distance (mm) | 250 | 175 | 350 | 130 |
| HAG conditions | ||||
| Glue | 10% PVA 4500, 5% ethylene glycol | 10% PVA 4500 | ||
| Optimized humidity (%) | 85 | 90 | ||
| Wavelength (Å) | 1.0000 | 1.0000 | 1.0000 | 1.0000 |
| Resolution range (Å) | 50.0–3.60 (3.66–3.60)a | 50.0–2.00 (2.03–2.00) | 50.0–3.40 (3.46–3.40) | 30–1.55 (1.58–1.55) |
| Space group | P212121 | P1 | P212121 | P21 |
| Unit-cell parameters | ||||
| a, b, c (Å) | 71.6, 133.0, 273.0 | 46.1, 59.2, 82.7 | 72.2, 133.4, 272.7 | 76.3, 53.2, 127.5 |
| α, β, γ (degrees) | 84.7, 90.2, 88.0 | 90, 93.6, 90 | ||
| No. of molecules/asymmetric unit | 1 | 2 | 1 | 2 |
| Total observations | 144,544 | 173,757 | 266,184 | 539,873 |
| Unique reflections | 30,118 | 56,452 | 37,120 | 143,400 |
| Completeness (%) | 95.6 (98.0) | 97.4 (96.1) | 99.8 (100) | 96.8 (94.4) |
| I/σ(I) | 21.4 (3.2) | 31.4 (5.6) | 36.9 (4.1) | 25.9 (3.2) |
| Rmerge | 0.042 (0.461) | 0.053 (0.157) | 0.046 (0.536) | 0.084 (0.420) |
| CC½ in outer shell (%) | 80.2 | 95.2 | 85.3 | 85.8 |
| Mosaicity (degrees) | 1.014 | 1.510 | 0.634 | 0.524 |
| Refinement | ||||
| Resolution range (Å) | 37.4–3.60 (3.72–3.60) | 35.7–2.01 (2.04–2.01) | 37.6–3.40 (3.49–3.40) | 29.9–1.55 (1.57–1.55) |
| Rwork/Rfree | 0.238/0.289 (0.286/0.338) | 0.201/0.261 (0.264/0.325) | 0.236/0.290 (0.271/0.327) | 0.167/0.196 (0.214/0.228) |
| No. of atoms | ||||
| Protein | 14,207 | 8012 | 14,222 | 8130 |
| Alginate | 61 | 122 | 48 | 72 |
| Water | 0 | 564 | 0 | 481 |
| Calcium ion | 0 | 6 | 0 | 2 |
| Chloride ion | 0 | 0 | 0 | 2 |
| PEG | 0 | 0 | 0 | 64 |
| RMSD | ||||
| Bond lengths (Å) | 0.003 | 0.010 | 0.003 | 0.007 |
| Bond angles (degrees) | 0.781 | 1.041 | 0.820 | 1.126 |
| Ramachandran plot | ||||
| Most favored (%) | 93.32 | 97.94 | 92.26 | 97.97 |
| Allowed (%) | 6.23 | 1.96 | 7.34 | 2.03 |
| Outlier (%) | 0.45 | 0.10 | 0.39 | 0 |
a Data in the highest-resolution shell are shown in parentheses.