Circular dichroism spectroscopy (CD) measurements were performed on a Jasco J-810 spectropolarimeter equipped with a Jasco CDF-426S Peltier set to 25°C. Averages of five scans were baseline-subtracted (PBS buffer; 25 mM phosphate, 150 mM NaCl) and the alpha-helical content was calculated from the molar ellipticity at 222 nm as previously described [16] (link).
For thermal stability experiments, spectra were obtained from 25°C to 80°C with 2.5°C increments. ApoA-I was diluted to 0.2 mg/ml in PBS (final concentration was 25 mM phosphate, 150 mM NaCl, pH 7.4), placed in a 1 mm quartz cuvette and, after extensive purging with nitrogen, scanned in the region 200 to 260 nm (scan speed was 20 nm/min). The Boltzmann function within the GraphPad software (GraphPad Software, Inc., CA, USA) was used to fit the molar ellipticity values at 222 nm of the temperature gradient to a sigmoidal fit curve.
For thermal stability experiments, spectra were obtained from 25°C to 80°C with 2.5°C increments. ApoA-I was diluted to 0.2 mg/ml in PBS (final concentration was 25 mM phosphate, 150 mM NaCl, pH 7.4), placed in a 1 mm quartz cuvette and, after extensive purging with nitrogen, scanned in the region 200 to 260 nm (scan speed was 20 nm/min). The Boltzmann function within the GraphPad software (GraphPad Software, Inc., CA, USA) was used to fit the molar ellipticity values at 222 nm of the temperature gradient to a sigmoidal fit curve.
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