Far UV CD Spectroscopy of Lymphostatin

The far UV CD spectrum of full-length lymphostatin (0.11 μm), rLifA^DTD/AAA (0.11 μm), and the digest fragment F1 (0.35 μm) were recorded at 10 nm/min; data pitch, 0.1 nm; response time, 2 s between 185 and 285 nm in a 0.1-cm path length quartz cuvette at 25 °C (JASCO-810 spectrometer). The proteins were exchanged into 10 mm sodium phosphate, pH 7.6, 150 mm sodium fluoride prior to analysis (HiTrap desalt column, GE) at 4 ml/min. Spectra were corrected by subtracting a buffer baseline, each an average of 5 spectra. Secondary structure was estimated using the Dichroweb CD secondary structure analysis server (20 (link)) including the methods CONTIN, SELCON3, and CDSSTR (21 (link)– (link)24 (link)) and reference data sets SP175 and 7 (25 (link)).