TABLE 3.
Analysis of stabilization of OLF variants by Ca2+
| Variant | Rationale | Tm | Tm + 10 mm CaCl2 | ΔTm |
|---|---|---|---|---|
| °C | °C | °C | ||
| myoc-OLF | Wild-type | 53.0 ± 0.5 | 59.6 ± 0.2 | 6.6 |
| myoc-OLF core | Identified structural corea | 49.7 ± 0.3 | 57.7 ± 0.1 | 8.1 |
| MBP-OLF(G246R) | Disease-causing variant | 42.5 ± 0.2 | 50.6 ± 0.0 | 8.1 |
| MBP-OLF(G252R) | Disease-causing variant | 43.0 ± 0.2 | 51.5 ± 0.2 | 8.5 |
| MBP-OLF(R272G) | Disease-causing variant | 41.0 ± 0.3 | 48.6 ± 0.1 | 7.6 |
| MBP-OLF(D273A) | Carboxylate side chain? | 21.7 ± 0.8 | 21.1 ± 0.7 | −0.6 |
| MBP-OLF(E323K) | Disease-causing variant | 44.0 ± 0.5 | 50.3 ± 0.2 | 6.2 |
| MBP-OLF(G364V) | Disease-causing variant | 45.0 ± 0.4 | 51.9 ± 0.1 | 6.9 |
| MBP-OLF(G367R) | Disease-causing variant | 42.7 ± 0.1 | 50.9 ± 0.4 | 8.1 |
| MBP-OLF(T377M) | Disease-causing variant | 44.3 ± 0.3 | 50.6 ± 0.6 | 6.3 |
| MBP-OLF(D378A) | Carboxylate side chain? | N/A | N/A | N/Ab |
| MBP-OLF(D380A) | Carboxylate side chain ligand? and disease causing variant | 46.6 ± 0.3 | 45.1 ± 0.5 | −1.5 |
| MBP-OLF(D384A) | Carboxylate side chain ligand? | 42.5 ± 0.7 | 53.6 ± 0.5 | 11.1 |
| MBP-OLF(E385A) | Carboxylate side chain ligand? | 39.7 ± 0.4 | 49.7 ± 0.3 | 10 |
| MBP-OLF(K423E) | Disease-causing variant | 34.2 ± 0.4 | 43.2 ± 0.1 | 9.0 |
| MBP-OLF(V426F) | Disease-causing variant | 41.5 ± 0.1 | 49.9 ± 0.1 | 8.4 |
| MBP-OLF(A427T) | Disease-causing variant | 48.3 ± 0.3 | 55.2 ± 0.4 | 6.9 |
| MBP-OLF(C433R) | disease causing variant | 40.4 ± 0.4 | 49.4 ± 0.5 | 9.0 |
| MBP-OLF(Y437H) | Disease-causing variant | 40.3 ± 0.4 | 48.6 ± 0.2 | 8.3 |
| MBP-OLF(I477N) | Disease-causing variant | 37.7 ± 0.8 | 46.8 ± 0.2 | 9.1 |
| MBP-OLF(I477S) | Disease-causing variant | 39.7 ± 0.2 | 48.2 ± 0.5 | 8.5 |
| MBP-OLF(N480K) | Disease-causing variant | 42.4 ± 0.2 | 48.1 ± 0.1 | 5.9 |
| MBP-OLF(P481L) | Disease-causing variant | 45.5 ± 0.4 | 51.2 ± 0.3 | 5.7 |
| MBP-OLF(I499F) | Disease-causing variant | 42.8 ± 0.1 | 50.4 ± 0.4 | 7.6 |
| MBP-OLF(S502P) | Disease-causing variant | 41.0 ± 0.3 | 49.9 ± 0.2 | 8.9 |
a From Ref. 27.
b Variant could not be purified in sufficient quantities in folded state for measurement.