Guidelines for Protocol Development:
Pressure-enhanced proteolytic digestion exploits the ability of high hydrostatic pressure to promote protein denaturation and the access of proteolytic enzymes to their target sites. Pressure denaturation is fundamentally different from thermal denaturation as it occurs by virtue of hydration of hydrophobic
residues and by water saturation of protein substrate cavities normally inaccessible to solvent. Pressure denaturation is more efficient for hydrophobic proteins, while some hydrophilic soluble proteins are reported to retain relatively compact conformation, even when saturated by water molecules [1-6].