Sweet and blind spots in E3 ligase ligand space revealed by a thermophoresis-based assay

December 4, 2020

Maiwald, S., Heim, C., Alvarez, B., et al.

ACS Medicinal Chemistry Letters 2020, doi: 10.1021/acsmedchemlett.0c00440


Repurposing E3 ubiquitin ligases for targeted protein degradation via customized molecular glues or proteolysis-targeting chimeras (PROTACs) is an increasingly important therapeutic modality. Currently, a major limitation in the design of suitable molecular glues and PROTACs is our fragmentary understanding of E3 ligases and their ligand space. We here describe a quantitative assay for the discovery and characterization of E3 ligase ligands that is based on the thermophoretic behavior of a custom reporter ligand. Thereby, it is orthogonal to commonly employed fluorescence-based assays and less affected by the optical properties of test compounds. It can be employed for the high-throughput screening of compound libraries for a given ligase but also for hit validation, which we demonstrate with the identification of unexpected well-binders and non-binders, yielding new insights into the ligand space of cereblon (CRBN).

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Topics: Monolith, MST, PROTACs, Publications

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