Table 1.
Crystallization of Rhodopsin
| Rhodopsin (17) | Rhodopsin (54) | Squid Rhodopsin (20) | Meta II (55) | Meta II (55) | Opsin (2) | Opsin (1) | |
|---|---|---|---|---|---|---|---|
| PDB ID | 1F88 | 1U19 | 2Z73 | 3PXO | 3PQR | 3CAP | 3DQB |
| Space Group | P41 | P41 | P62 | H32 | H32 | H3 | H32 |
| Resolution | 2.8Å | 2.2Å | 2.5Å | 3.0Å | 2.85Å | 2.9Å | 3.2Å |
| Method | Hanging Drop | Hanging Drop | Sitting Drop | Hanging Drop | Hanging Drop | Hanging Drop | Hanging Drop |
| Protein Concentration | 10 mg/mL | 6–8 mg/mL | 8 mg/mL | 5 mg/mL | 5 mg/mL | 5 mg/mL | 7–10 mg/mL |
| Drop Size | 9–11 μL | 4–10 μL | 10 μL | 4μL | 4μL | 4μL | 4μL |
| Source | Bovine ROS | Bovine ROS | Microvillar membranes of photoreceptor rhabdoms from squid retina | Bovine ROS | Bovine ROS | Bovine ROS | Bovine ROS |
| Protein Buffer | MES or sodium acetate pH 6.3–6.4, 5–7 mM βME, 90–120 mM Zn(OAc)2, 0.50–0.65% heptane-1,2,3-triol, 2.2:1 ratio of nonyl glucoside: rhodopsin | 6–12 mM βME, 0.1–0.5% heptyl- thioglucoside, 0.5–0.7 M ammonium sulfate | 30 mM MES, pH 6.4, 30 mM Zn(OAc)2, ~100 mM β-D-octyl- glucopyranoside | 1% β-D-octyl- glucopyranoside, 0.02% n- dodecyl- β-D- maltopyranoside | 1% β-D-octyl- glucopyranoside, 0.02% n- dodecyl- β-D- maltopyranoside, GαCT2 peptide | 1% β-D-octyl- glucopyranoside | 1% β-D-octyl- glucopyranoside, GαCT peptide (4:1 ratio of protein:peptide) |
| Reservoir Solution | MES-buffered 3.0–3.4 M ammonium sulfate pH 6.0–6.1 | 20–30 mM MES pH 5.9–6.1, 2.5–3.0 M ammonium sulfate | 3.2 M ammonium sulfate, 32 mM MES pH 6.0–6.7, 38 mM EDTA, 10 mM βME | 3.0–3.4 M (NH4)2SO4, 0.1 M sodium acetate pH 5.0–5.8 | 3.0–3.4 M (NH4)2SO4, 0.1 M sodium acetate pH 5.0–5.8 | 2.8–3.2 M ammonium sulfate, 0.1 M MES or sodium acetate pH 5.6 | 3.2 M ammonium sulfate, 0.1 M MES or sodium citrate pH 6.0 |
| Drop Size | 3–5°C | 10°C | 4°C | 4°C | 4°C | 4°C | 4°C |