Table 1.
Variant | Tm (K) | ΔHm (kJ mol−1) | ΔCp (kJ mol−1 K−1) | m (kJ mol−1 M−1) | ΔGf (kJ mol−1) | [D]50% (M) | ΔΔGf (kJ mol−1) |
---|---|---|---|---|---|---|---|
aI30K, I57A | 327.4 ± 3.1 | −99 ± 11 | −0.4 ± 0.8 | 8.8 ± 1.8 | −8.2 ± 0.6 | 0.9 ± 0.2 | |
A16D, P33Q, G35E, V38M, Q59K | 333.3 ± 1.9 | −134 ± 9 | −2.6 ± 0.3 | 5.4 ± 0.5 | −9.1 ± 0.6 | 1.7 ± 0.2 | |
K18E, P33Q, V47D | 336.0 ± 1.7 | −147 ± 10 | −2.9 ± 0.1 | 4.3 ± 0.2 | −10.1 ± 1.0 | 2.3 ± 0.2 | |
aI44N, I57A | 339.0 ± 2.4 | −163 ± 28 | −2.6 ± 0.4 | 7.4 ± 1.5 | −13.0 ± 2.8 | 1.8 ± 0.5 | |
A27V, D52G | 341.7 ± 1.0 | −211 ± 19 | −3.6 ± 0.3 | 6.0 ± 0.8 | −16.4 ± 1.5 | 2.7 ± 0.4 | |
aE41G, F50L, I57A | 331.6 ± 0.5 | −191 ± 3 | −1.5 ± 0.1 | 11.5 ± 0.2 | −16.6 ± 0.4 | 1.4 ± 0.0 | |
A16T, N56I | 339.4 ± 0.6 | −227 ± 19 | −4.1 ± 0.3 | 7.0 ± 0.4 | −17.0 ± 1.5 | 2.4 ± 0.2 | |
aV31D, I57A | 344.7 ± 0.5 | −222 ± 0 | −3.7 ± 0.0 | 7.0 ± 0.0 | −17.7 ± 0.1 | 2.5 ± 0.0 | |
aI57A | 333.1 ± 0.5 | −269 ± 14 | −5.4 ± 0.5 | 11.6 ± 0.6 | −17.9 ± 0.6 | 1.5 ± 0.1 | |
G10E, T39S | 339.7 ± 0.7 | −238 ± 16 | −4.0 ± 0.7 | 8.5 ± 0.5 | −18.4 ± 0.4 | 2.2 ± 0.1 | |
aM40T, R48S, I57V | 344.3 ± 0.3 | −235 ± 9 | −3.8 ± 0.2 | 7.1 ± 0.1 | −19.1 ± 0.4 | 2.7 ± 0.1 | |
G35W, A58V | 336.8 ± 0.1 | −281 ± 19 | −5.7 ± 0.5 | 6.9 ± 0.2 | −19.1 ± 0.9 | 2.8 ± 0.1 | |
E4G | 341.5 ± 0.1 | −259 ± 7 | −4.0 ± 0.2 | 7.3 ± 0.1 | −21.4 ± 0.3 | 2.9 ± 0.1 | |
L32Q, I57S | 348.1 ± 0.6 | −275 ± 5 | −4.4 ± 0.0 | 6.1 ± 0.2 | −22.9 ± 0.7 | 3.8 ± 0.2 | |
V13E | 343.1 ± 0.2 | −280 ± 23 | −4.1 ± 0.5 | 9.6 ± 0.6 | −24.0 ± 1.6 | 2.5 ± 0.2 | |
aE15G, M40T, R48C, I57V | 347.6 ± 0.1 | −281 ± 8 | −4.1 ± 0.1 | 8.3 ± 0.2 | −24.9 ± 0.7 | 3.0 ± 0.1 | |
aV38E, I57V | 348.8 ± 0.3 | −295 ± 16 | −4.4 ± 0.3 | 8.9 ± 0.5 | −25.8 ± 1.2 | 2.9 ± 0.2 | |
T39A, I57N | 343.5 ± 0.2 | −312 ± 0 | −4.7 ± 0.0 | 10.6 ± 0.1 | −26.4 ± 0.1 | 2.5 ± 0.0 | |
L49I | 350.3 ± 0.2 | −322 ± 8 | −4.6 ± 0.1 | 8.7 ± 0.2 | −28.9 ± 0.8 | 3.3 ± 0.1 | 3.4 ± 0.1 |
V34E, D45Y | 344.3 ± 0.0 | −363 ± 7 | −6.0 ± 0.2 | 8.4 ± 0.2 | −29.2 ± 0.5 | 3.5 ± 0.1 | |
wild-type | 352.3 ± 0.2 | −322 ± 11 | −4.3 ± 0.2 | 8.2 ± 0.1 | −30.5 ± 0.8 | 3.7 ± 0.1 | 0 |
P33L | 354.1 ± 0.5 | −330 ± 43 | −4.2 ± 0.7 | 8.3 ± 0.8 | −32.5 ± 3.5 | 3.9 ± 0.6 | |
aL49I, I57V | 356.2 ± 0.3 | −332 ± 13 | −4.2 ± 0.2 | 7.9 ± 0.4 | −32.9 ± 1.3 | 4.2 ± 0.3 | −3.8 ± 0.1 |
aI57V | 354.5 ± 0.2 | −349 ± 12 | −4.7 ± 0.2 | 8.3 ± 0.2 | −33.3 ± 0.9 | 4.0 ± 0.1 | −2.2 ± 0.1 |
aD55G, I57V | 361.6 ± 0.3 | −364 ± 13 | −4.3 ± 0.2 | 8.5 ± 0.2 | −38.1 ± 0.8 | 4.5 ± 0.1 | −6.5 ± 0.2 |
D55G | 361.1 ± 0.1 | −378 ± 10 | −4.7 ± 0.1 | 8.5 ± 0.2 | −38.4 ± 1.0 | 4.5 ± 0.2 | −6.9 ± 0.3 |
aVariant selected from the I57A library.
Tm, is the melting temperature in the absence of denaturant. ΔHm is the enthalpy change for folding at Tm. ΔCp is the change in heat capacity for folding. m is the m-value for GuHCl unfolding. ΔGf is the free energy for folding at 298 K. [D]50% is the midpoint for the GuHCl unfolding at 298 K and ΔΔGf is the change in stability at 298 K relative to wild-type from a fit of six variants with a common m-value of 8.4 ± 0.3 kJ mol−1 M−1. The errors are the propagated standard errors from the global fits of the data.