Mechanisms of protein evolution have been of much interest in the peptide and protein fields. One such mechanism, discovered recently, involves peptides evolving within a “host” protein and emerging via proteolytic processing. The first examples of this mechanism stem from peptides buried within the precursors of seed storage albumins, resulting in what has now been established as the PawS Derived Peptide (PDP) family¹. Indications also suggested peptides can emerge from the precursor protein of another class of seed storage proteins in the same fashion, and recent work has now established another peptide family, the Vicilin Buried Peptide (VBP) family². To date limited structural work has been conducted on the VBP family. We here present an extensive structural characterization of members of the VBP family to better understand their structural features. Six members of the VBP family were synthesized by solid phase peptide synthesis and subjected to NMR spectroscopy, with four full 3D structures being determined. Our work confirms that the familial structure of the VBP family is the helix-loop-helix hairpin fold stapled by disulfide bonds in a 1-4, 2-3 ladder like fashion. In addition, we establish that numerous interhelix interactions, in the form of hydrophobic contacts and/or salt bridges, contribute to the stability of the fold and control the angle at which the two anti-parallel helices interface. Reported functions for VBPs include ribosomal inhibition and trypsin inhibition. Given the age and evolutionary retention, combined with conserved structural features and different bioactivities, leads us to believe that these peptides play an important role in plant biology.