Proteolysis-PrEP: Optimization of Pressure Cycling Conditions for Protein Digestion with Trypsin in Solution
Pressure Cycling Technology (PCT) has been proven to accelerate enzymatic protein digestion. The positive effect of PCT on trypsin digestion has been demonstrated by several laboratories [1, 2, 3] who have reported improved sequence coverage, higher recovery and significantly reduced digestion times when the trypsin reactions were carried out under pressure. Not only has PCT been shown to accelerate and improve digestion in solution, but it can also accelerate in-gel trypsin digestion [4, 5]. Additionally, the enhancing effect of PCT on the activity of several other enzymes, including Proteinase K, PNGase F, Lys-C and lysozyme has been reported [6, 7, 8, 9].