Development of an Efficient Enzyme Production and Structure-Based Discovery Platform for BACE1 Inhibitors.

ABSTRACT: BACE1 (Beta-site Amyloid Precursor Protein (APP) Cleaving Enzyme 1) is a promising therapeutic target for Alzheimer's Disease (AD). However, efficient expression, purification, and crystallization systems are not well described or detailed in the literature nor are approaches for treatment of enzyme kinetic data for potent inhibitors well described. We therefore developed a platform for expression and purification of BACE1, including protein refolding from E.coli inclusion bodies, in addition to optimizing a reproducible crystallization procedure of BACE1 bound with inhibitors. We also report a detailed approach to the proper analysis of enzyme kinetic data for compounds that exhibit either rapid-equilibrium or tight-binding mechanisms. Our methods allow for the purification of ∼15

SUBMITTER: Yen YC 

PROVIDER: S-EPMC7284891 | biostudies-literature | 2019 Nov

REPOSITORIES: biostudies-literature