Table 1.
Thermal denaturation
MelBStBufferaNumber of tests10% unfolding (°C)90% unfolding (°C)50% unfolding (Tm, °C)ΔTm (°C; P value)
WT Apo 46.13 ± 0.70b 62.26 ± 0.50 53.29c ± 0.05 
K+ 46.06 ± 0.02 59.94 ± 0.69 53.34 ± 0.04 0.05d (>0.05e
Na+ 49.07 ± 0.28 60.06 ± 0.29 54.83 ± 0.03 1.54 (<0.05) 
Mel 45.95 ± 0.47 64.24 ± 0.42 54.07 ± 0.07 0.78 (<0.05) 
Mel and Na+ 49.42 ± 0.13 61.76 ± 0.09 56.71 ± 0.05 3.43 (<0.05) 
D55C Apo   >80f  
K+ 46.51 ± 0.71 65.07 ± 0.06 56.15 ± 0.16 >14 
Na+ 46.35 ± 0.25 66.51 ± 0.31 56.75 ± 0.25 >14 
Mel   >80  
Mel and Na+ 46.25 ± 0.20 68.52 ± 0.27 58.15 ± 0.11 1.40 (<0.05) 
MelBStBufferaNumber of tests10% unfolding (°C)90% unfolding (°C)50% unfolding (Tm, °C)ΔTm (°C; P value)
WT Apo 46.13 ± 0.70b 62.26 ± 0.50 53.29c ± 0.05 
K+ 46.06 ± 0.02 59.94 ± 0.69 53.34 ± 0.04 0.05d (>0.05e
Na+ 49.07 ± 0.28 60.06 ± 0.29 54.83 ± 0.03 1.54 (<0.05) 
Mel 45.95 ± 0.47 64.24 ± 0.42 54.07 ± 0.07 0.78 (<0.05) 
Mel and Na+ 49.42 ± 0.13 61.76 ± 0.09 56.71 ± 0.05 3.43 (<0.05) 
D55C Apo   >80f  
K+ 46.51 ± 0.71 65.07 ± 0.06 56.15 ± 0.16 >14 
Na+ 46.35 ± 0.25 66.51 ± 0.31 56.75 ± 0.25 >14 
Mel   >80  
Mel and Na+ 46.25 ± 0.20 68.52 ± 0.27 58.15 ± 0.11 1.40 (<0.05) 
a

CD assay buffers consisted of 10 mM KPi, pH 7.5, 10% glycerol, and 0.035% UDM with a given component at 50 mM of each.

b

SEM.

c

Tm value at which 50% protein unfolded.

d

The difference in Tm value when compared with apo (in WT) or the buffer containing Na+ (in D55C mutant).

e

Unpaired t test; P < 0.05 was considered statistically significant.

f

Completion of denaturation cannot be approached, so this value is an estimation based on the unfolded fraction.

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