Table 1.
Differential scanning calorimetric data of Fc-wt and all Fc variants that were expressed solubly in P. pastoris. Depicted are the temperatures of unfolding (Tm) and the differences in the Tm values compared to Fc-wt (ΔTm). The first transition represents the unfolding process of the CH2 and the second and third the ones of the CH3-domain. Averages and standard deviations from three experiments are shown.
| Variant | Tm1 (°C) | Tm2 (°C) | Tm3 (°C) | ΔTm1 | ΔTm2 | ΔTm3 |
|---|---|---|---|---|---|---|
| Fc-wt | 65.7 ± 0.1 | 78.1 ± 0.1 | 82.6 ± 0.0 | – | – | – |
| V263A | 63.1 ± 0.1 | 77.5 ± 0.0 | 82.2 ± 0.0 | − 2.6 | − 0.6 | − 0.4 |
| Q347E | 64.9 ± 0.1 | 81.6 ± 0.1 | 86.4 ± 0.0 | − 0.8 | 3.5 | 3.8 |
| S354P | 65.1 ± 0.2 | 80.0 ± 0.1 | 85.8 ± 0.1 | − 0.6 | 1.9 | 3.2 |
| K360E | 64.9 ± 0.2 | 80.4 ± 0.1 | 85.1 ± 0.1 | − 0.8 | 2.3 | 2.5 |
| K360Q | 65.4 ± 0.0 | 81.6 ± 0.1 | 85.7 ± 0.0 | − 0.3 | 3.4 | 3.1 |
| K392R | 64.7 ± 0.1 | 79.1 ± 0.1 | 84.4 ± 0.1 | − 1.0 | 0.9 | 1.8 |
| S400F | 65.3 ± 0.1 | 80.1 ± 0.2 | 85.4 ± 0.1 | − 0.4 | 2.0 | 2.8 |
| Q418L | 65.0 ± 0.1 | 81.0 ± 0.1 | 86.3 ± 0.1 | − 0.7 | 2.9 | 3.7 |
| N421D | 65.4 ± 0.4 | 79.5 ± 0.1 | 84.2 ± 0.0 | − 0.3 | 1.4 | 1.6 |
| S424T | 65.2 ± 0.4 | 81.0 ± 0.1 | 85.9 ± 0.1 | − 0.5 | 2.9 | 3.3 |
| T437I | 64.4 ± 0.1 | 80.2 ± 0.1 | 84.3 ± 0.0 | − 1.3 | 2.0 | 1.7 |
| Q438K | 64.0 ± 0.1 | 80.4 ± 0.1 | 85.2 ± 0.1 | − 1.7 | 2.3 | 2.6 |
| Q438R | 64.1 ± 0.1 | 79.5 ± 0.1 | 83.9 ± 0.0 | − 1.6 | 1.4 | 1.3 |
| Q347E, Q418L | 64.9 ± 0.1 | 85.0 ± 0.0 | 90.2 ± 0.0 | − 0.8 | 6.9 | 7.6 |
| K360Q, Q418L | 64.9 ± 0.1 | 84.6 ± 0.1 | 89.6 ± 0.1 | − 0.8 | 6.5 | 7.0 |
| K360Q Q438R | 63.7 ± 0.1 | 82.8 ± 0.1 | 87.2 ± 0.0 | − 2.0 | 4.7 | 4.6 |
| K360Q, Q418L, Q438R | 63.8 ± 0.0 | 86.5 ± 0.1 | 91.0 ± 0.1 | − 1.9 | 8.4 | 8.4 |
| K360Q, G446C | 64.7 ± 0.3 | 83.2 ± 0.1 | 90.4 ± 0.1 | − 1.0 | 5.0 | 7.8 |