Synthesis of substrate peptides incorporating non-natural amino acids and screening study using BACE1

Peptide library screening is used to detect optimal sequences for enzymatic cleavage; moreover, the data obtained through this screening are useful for the establishment of a fast screening system and designing of substrate-based enzyme inhibitors. In this study, peptide libraries were prepared and digested with the beta-site amyloid precursor protein cleaving enzyme (BACE1). BACE1 has been used as a target enzyme for drug development against Alzheimer's disease (AD). The library sequences were derived from our previous screening study based on amyloid-beta precursor protein (APP) substrates. Then, newly selected non-natural amino acids were incorporated into several positions on these sequences. After digestion with BACE1, the reaction mixtures were analyzed with high-performance liquid chromatography followed by mass spectrometry to identify the peptides undergoing efficient cleavage. The data obtained from this study can be used for designing drugs against AD in the future.