Cdf 426s 15 peltier temperature control system

Circular dichroism (CD) spectra were acquired on a JASCO J-815 CD spectrometer controlled by a CDF-426S/15 Peltier temperature control system (Jasco Analytical Instruments, Easton, MD, USA) using a quartz cuvette with a 1-cm path length. The enzyme was prepared in phosphate buffer (20 mM sodium phosphate, 150 mM NaCl, pH 7.5) at a final concentration of 8 μM. All spectra were obtained at 20°C in the range 200–260 nm with a bandwidth of 2 nm and a response time of 4 s/nm. CD spectra were buffer subtracted and normalized to mean residue ellipticity. Thermal unfolding experiments were monitored at 222 nm in the temperature range 20–90°C with a scan rate of 1°C.min^-1. The melting temperature (T_M) was determined according to the sigmoidal-Boltzmann fitting of the CD denaturation curve.

Circular Dichroism spectroscopy was employed to assess the global conformation of EaBglA in both tetrameric and monomeric conformations. CD measurements were acquired on a JASCO J-815 CD spectrometer controlled by a CDF-426S/15 Peltier temperature control system (Jasco Analytical Instruments, Oklahoma, EUA). A quartz cuvette with a 1-cm path length was used for all CD experiments and each spectrum was an average of at least three scans. Protein concentration was 5.8 μM (for tetramer and monomer) in 20 mM sodium phosphate, 150 mM NaCl, pH 7.5. All spectra were obtained at 20 °C in the range 200–260 nm with a bandwidth of 2 nm and a response time of 4 s/nm. CD data were buffer subtracted and normalized to molar residual ellipticity allowing the comparison between the forms.