Applying Ht-Saxs to Chemical Ligand Screening

Authors

Chris A Brosey
Runze Shen
Davide Moiani
Darin E Jones
Kathryn Burnett
Greg L Hura
John A Tainer

Publication Date

1-1-2023

Journal

Methods in Enzymology

Abstract

Chemical probes are invaluable tools for investigating essential biological processes. Understanding how small-molecule probes engage biomolecular conformations is critical to developing their functional selectivity. High-throughput solution X-ray scattering is well-positioned to profile target-ligand complexes during probe development, bringing conformational insight and selection to traditional ligand binding assays. Access to high-quality synchrotron SAXS datasets and high-throughput data analysis now allows routine academic users to incorporate conformational information into small-molecule development pipelines. Here we describe a general approach for benchmarking and preparing HT-SAXS chemical screens from small fragment libraries. Using the allosteric oxidoreductase Apoptosis-Inducing Factor (AIF) as an exemplary system, we illustrate how HT-SAXS efficiently identifies an allosteric candidate among hits of a microscale thermophoresis ligand screen. We discuss considerations for pursuing HT-SAXS chemical screening with other systems of interest and reflect on advances to extend screening throughput and sensitivity.

Keywords

X-Ray Diffraction, Ligands, Scattering, Small Angle, Oxidoreductases, Synchrotrons, IF, Allostery, Chemical screening, Drug discovery, Fragment libraries, HT-SAXS, Redox, X-ray scattering

DOI

10.1016/bs.mie.2022.09.022

PMID

36641213

PMCID

PMC11239221

PubMedCentral® Posted Date

7-11-2024

PubMedCentral® Full Text Version

Author MSS

Published Open-Access

yes