Tm

Stability/Folding

Melting Temperature (Tm)

°C (Celsius)

Circular Dichroism (CD)

50 mM sodium phosphate

6.5

50 μM

218 nm

Tm = melting temperature
CD = circular dichroism, RC = random coil
The CD data were collected on Jasco-810 spectrometer equipped with a thermoelectric unit and using a 0.1 mm path-length cell. Protein samples were 50 μM in 50 mM sodium phosphate at pH 6.5. Thermal melts were monitored at 218 nm. Data were collected every 1 deg.C with an equilibration time of 2 min. Far-UV spectra were acquired in the continuous mode at 25 °C with 1 nm band- width and a 4 s response time. For the thermal denatura- tion curves, the data were normalized by first shifting all points linearly, such that the [t]218 value at 5 °C was zero. Then a scaling factor was obtained for each set by dividing the maximum [t]218 value for all sets at 95 °C (i.e. 53.0) by the [t]218 value at 95 °C for each set. All data points for each set were then scaled by the unique scaling factor calculated for each set.