Figure 3
Trypsin digestion analysis and ATR-FTIR spectroscopy of HomA and HomB. (A) pictorial representation of HomA and HomB protease digestion of scheme. (B) trypsin digested proteins run onto denaturing SDS-PAGE. (C) same protein sample run onto the semi-native gel with and without boiling. (D) percentage of folded and unfolded fractions observed on semi-native PAGE. The red asterisk (*) denotes the N-terminal part that comes out upon unfolding. (E–H) Secondary structure analysis using curve fitting of the amide I region (1600–1700 cm−1) ATR-FTIR spectroscopy. (E,F) FTIR absorbance spectra of amide 1 of HomA and HomB, respectively, with their second derivatives. (G,H) Deconvolution of amide 1 spectra of HomA and HomB, respectively, for secondary structure content analysis. The spectral range of 1600–1700 cm−1 was decomposed using Origin 8.0 software. [Full gel image for (B) and (C) are in Supplementary Fig. S8].
