Structural Biology & Biophysics Core (SBB)

PROJECT SUMMARY/ABSTRACT Structural Biology and Biophysical (SBB) Core The Structural Biology & Biophysics (SBB) Core of the Indiana University School of Medicine Purdue University TaRget Enablement to Accelerate Therapy Development for Alzheimer's Disease (IUSM-Purdue TREAT-AD) Center will help develop high quality lead compounds for novel Alzheimer's Disease (AD) targets. The SBB Core will work in concert with the other Technical Cores to select, produce, characterize, and structurally enable AD targets in support of structure-based drug design for lead optimization. The SBB Core will utilize a two-pronged approach to support the overall goals of the TREAT-AD center. The SBB Core will first characterize the biochemical and biophysical properties of target proteins in the unbound and ligand-bound forms to better understand the kinetics and thermodynamics of ligand engagement with the target. Enzyme kinetic studies, including the use of advanced Desorption Electrospray Ionization Mass Spectrometry (DESI-MS) assays, will be used to determine steady-state kinetic parameters for substrates and ligands. Surface Plasmon Resonance (SPR) will be used to determine ligand-binding kinetics and Isothermal Titration Calorimetry (ITC) and MicroScale Thermophoresis (MST) will be used to determine ligand-binding thermodynamics. Mass Photometry (MP) and size-exclusion chromatography (SEC) with Multi-Angle Light Scattering (MALS) will be used to determine protein quaternary structure and the influence of ligand binding on oligomeric state. Second, the SBB core will work to determine the structures for prioritized targets via X-ray diffraction and/or cryo-electron microscopy. The SBB core will develop approaches and integrated workflow in close collaboration with the MCCB and ADHTS cores for each target protein to acquire data in support SAR analysis and structure-based drug design. The SBB core will also help establish connectivity between molecular design and biological function to enhance understanding of the role of the target proteins in AD processes. The SBB core will work to accomplish these goals and those of the IUSM-Purdue TREAT-AD center through the following Specific Aims: 1) Evaluate the feasibility of nominated protein targets for structure-based design studies, 2) Produce and characterize the biochemical, biophysical, and structural properties of target proteins. 3) Elucidate target structures to guide drug discovery activities.