Figure 3

Model of Glo-3 endoproteolytic processing. The observed M_r, pI, MS/MS sequence data, and N-terminal sequence data from this study and previous studies [26, 58] were reconciled with theoretical peptides created by the endoproteolytic cleavage of preproglobulin-3 (GenBank Accession JQ954759). A linear representation of preproglobulin-3 is shown with approximate locations of potential cleavage sites, labeled 1–4. The protein domains are represented as follows: signal sequence (SS) (white); N-terminal sequence (grey); vicilin domain (divided into three segments by cleavage sites – N-terminal segment (green), middle segment (blue), C-terminal segment (orange). Red boxes correspond to the location of the linear epitopes used when creating the anti-Glo-3A polyclonal antibodies. Black boxes correspond to the location of MS/MS sequenced peptides. Purple boxes correspond to sequence obtained from N-terminal sequencing. The observed and expected molecular masses (kDa) and isoelectric points (pI) of the resulting polypeptides following endoproteolytic cleavage is indicated on the right. Additionally, the expected processing of proteins inferred by previous studies (Dupont et al., 2011 [20] and Singh et al., 2001 [26] have been demonstrated.