Denaturation/renaturation profile of MpNep2.

<p><b>A</b>. Thermal denaturation plot of MpNep2 monomer monitoring 222 nm absorbance by circular dichroism. Right and left insets show far-UV scans from 260 to 210 nm obtained during the heating and cooling experiment, respectively. <b>B</b>. Thermal denaturation plot of MpNep2 monomer monitoring the center of spectral mass of the Trp fluorescence emission during the heating (black filled circles) and cooling (black open circles). Open triangle shows the center of mass after 1 h of cooling. Light scattering during heating (blue filled circle) and cooling (blue open circles) were also recorded. Inset shows urea reversibility plot monitoring center of mass of the Trp fluorescence emission. For thermal denaturation, proteins were left for 10 min in each temperature before recording the Trp spectra. <b>C</b>. Relative Trp fluorescence emission spectrum at 20°C, 40°C, 58°C and 1 h after cooling to 20°C. Experiments are shown as mean (n = 3) with independent protein preparatoins.