Circular dichroism spectra of the CT domain in solution. CD spectra of the CT domain at three different pH conditions: a) pH 4, b) pH 8 and c) pH 12, all display alpha-helical patterns. The α-helical proteins commonly show negative bands at 222 nm and 208 nm minima and a positive band at 193 nm). 


METHODS
Samples with concentrations of 59-65 µM (0.18-0.2 mg/ml) for the helix No4 peptide and 0.2 mg/ml for the full-length CT domain were used for the CD experiment. The peptides and CT domain were analyzed with a Chirascan CD spectrometer (Applied Photophysics) and a 1 mm quartz cuvette. The temperature was maintained at 20 °C. The wavelength scan range was 190−260 nm (1 nm steps), the bandwidth was 1 nm, and the time per point was 1 s, with triplicate measurements for each sample. Data was smoothed using Savitzky-Golay filter with 7 points of window. The deconvolution analysis of CT-domain in solution was performed using the BeStSel software.


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