(A) Location of potential trypsin cleavage sites (Lys and Arg residues) in the CHP3 sequence. The two major sites, at which the cleavage is effective in the limited proteolysis are highlighted in red. (B) The full-size gels of limited proteolysis in different conditions. Proteins samples are indicated on the left, the ion conditions – on the top, trypsin only (T), or different CHP3 samples (CHP3, mCHP3, CHP3:CBD, or mCHP3:CBD) incubated with trypsin for 0, 5, 15, 30, 45, and 60 min were loaded from the left to the right. For a band annotation, see Figure 4A. Gels of CHP3 (MgCl2+CaCl2) and of CHP3:CBD (MgCl2+CaCl2) from Figure 4A (C) LC–ESI-TOF mass spectrometry analysis of major proteolytic fragments (1 and 2), the detected masses were 17,922 Da for the fragment 1 and 9630 Da for the fragment 2.