6NV9

BACE1 in complex with a macrocyclic inhibitor

Summary for 6NV9

• Entry DOI: 10.2210/pdb6nv9/pdb
• Descriptor: Beta-secretase 1, (3S)-3-hydroxy-N-(2-methylpropyl)-N~2~-{[(4S)-17-[(methylsulfonyl)(propyl)amino]-2-oxo-3-azatricyclo[13.3.1.1~6,10~]icosa-1(19),6(20),7,9,15,17-hexaen-4-yl]methyl}-L-norleucinamide, SULFATE ION, ... (4 entities in total)
• Functional Keywords: protease, inhibitor complex, peptide binding protein
• Biological source: Homo sapiens (Human)
• Total number of polymer chains: 3
• Total formula weight: 132452.67

Authors

Yen, Y.C.,Ghosh, A.K.,Mesecar, A.D. (deposition date: 2019-02-04, release date: 2019-10-09, Last modification date: 2024-11-13)

Primary citation

Yen, Y.C.,Kammeyer, A.M.,Jensen, K.C.,Tirlangi, J.,Ghosh, A.K.,Mesecar, A.D.
Development of an Efficient Enzyme Production and Structure-Based Discovery Platform for BACE1 Inhibitors.
Biochemistry, 58:4424-4435, 2019

PubMed 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 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 mg of BACE1 enzyme from 1 L of culture which is higher than reported yields in the current literature. To evaluate the data analysis approach developed here, a well-known potent inhibitor and two of its derivatives were tested, analyzed, and compared. The inhibitory constants () obtained from the kinetic studies are in agreement with dissociation constants () that were also determined using isothermal titration calorimetry (ITC) experiments. The X-ray structures of these three compounds in complex with BACE1 were readily obtained and provide important insight into the structure and thermodynamics of the BACE1-inhibitor interactions.

PubMed: 31549827
DOI: 10.1021/acs.biochem.9b00714

Experimental method

X-RAY DIFFRACTION (2.13 Å)