Far-UV CD Spectroscopy of Cyp40 Protein

The far UV CD spectrum of 0.75 μM Cyp40 was recorded at 20 nm min⁻¹; data pitch 0.1 nm; response time 1 s between 185 and 285 nm in a 0.1 cm path-length quartz cuvette between 10°C and 60°C at 10°C intervals (JASCO-810 spectrometer). The protein was exchanged into 10 mM sodium phosphate, pH 8; 150 mM sodium fluoride prior to analysis (HiTrap desalt column, GE). Spectra were corrected by subtracting a buffer baseline recorded at each temperature, spectra were recorded in triplicate. Secondary structure was estimated using the Dichroweb CD secondary structure analysis server using the methods CONTIN, SELCON3 and CDSSTR and data set SP175; mean results are presented with S.E.M [29 (link)–34 (link)].