Work

Ligand Screening and Discovery Using Cocktail Soaking and Automated Microcrystal Electron Diffraction

Year: 2025

Type: article

Abstract: Cocktail soaking using single-crystal X-ray diffraction previously allowed high-throughput crystallographic screening of ligands against protein targets. However, protein microcrystals are not amenable to this approach if they are too small to yield strong diffraction patterns. Herein, a workflow integrating cocktail soaking with automated microcrystal electron diffraction to allow rapid ligand screening, structure determination, and binding analysis directly from microcrystals is developed. This can improve the successful hit rate, because binding is often more efficient when smaller crystals are soaked in the ligand. The approach is validated with known ligands of thermolysin and identified novel binding interactions for ligands of proteinase K. The structures of multiple protein-ligand complexes, including ligands with weak binding affinities, can be solved rapidly. Their estimated relative binding affinities are in good agreement with previous work and independent microscale thermophoresis measurements.

Source: ChemMedChem

DOI: 10.1002/cmdc.202500156

Authors: Jieye Lin, Marc J. Gallenito, Johan Hattne, Tamir Gonen

Institutions: University of California, Los Angeles, Howard Hughes Medical Institute

Language: English

Topic: Enzyme Structure and Function

Subfield: Materials Chemistry

Field: Materials Science

Domain: Physical Sciences

Open Access status: hybrid

Estimated APC paid: $4,150

Funders: U.S. Department of Defense, Howard Hughes Medical Institute, National Institutes of Health, National Institute of General Medical Sciences