Denaturation-sensitive BRET probes enable intracellular thermal shift measurements across diverse protein targets and biomolecular complexes

Denaturation-sensitive BRETSA in living cells measures thermal shift profiles for over 500 globular proteins across 24 target families, enabling longitudinal tracking with luciferase-tagged targets.

• Why it matters: Thermal shift assays can be constrained by sensitivity and scalability, limiting how broadly and repeatedly target engagement can be assessed during drug discovery.
• What they did: The authors combine DSF and CETSA concepts to create BRETSA, using denaturation-sensitive BRET probes with luciferase-tagged proteins to record temperature-dependent denaturation in cells.
• The result: Target engagement is confirmed for over 200 proteins, and BRETSA extends to biomolecular complexes to probe cooperative binding for molecular glues and tricomplex drug modalities.

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