SUMMARY

•  HiPOS enables real time analysis of enzymes at highpressure for rapid optimization of reaction conditions

•  HiPOS is a useful tool for monitoring protein unfoldingand refolding where proper refolding restores activity

•  ALP activity persists in 4 M urea, the conditionstypically used for lysyl endopeptidase (Lys C) digestionof proteins, allowing the opportunity for spuriousdephosphorylation of protein analytes ALP isinactivated at the same elevated pressure andtemperature at which Lys C activity is accelerated

•  ALP activity persists in 0 8 M urea, the conditionstypically used trypsin digestion of proteins ALP activityis decreased at the same pressure at which trypsindigestion is enhanced