High-throughput peptide-centric local stability assay extends protein-ligand identification to membrane proteins, tissues and bacteria
Systematic mapping of protein-ligand interactions is essential for understanding biological processes and drug mechanisms. Peptide-centric local stability assay (PELSA) is a powerful tool for detecting these interactions and identifying potential binding sites. However, its original workflow is limi...
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| Main Authors: | , , , , , , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
2026
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| In: |
Nature structural & molecular biology
Year: 2026, Volume: 33, Issue: 1, Pages: 184-192 |
| ISSN: | 1545-9985 |
| DOI: | 10.1038/s41594-025-01699-y |
| Online Access: | Verlag, kostenfrei, Volltext: https://doi.org/10.1038/s41594-025-01699-y Verlag, kostenfrei, Volltext: https://www.nature.com/articles/s41594-025-01699-y 
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| Author Notes: | Kejia Li, Clement M. Potel, Isabelle Becher, Nico Hüttmann, Martin Garrido-Rodriguez, Jennifer Schwarz, Mira Lea Burtscher, Mikhail M. Savitski |
| Summary: | Systematic mapping of protein-ligand interactions is essential for understanding biological processes and drug mechanisms. Peptide-centric local stability assay (PELSA) is a powerful tool for detecting these interactions and identifying potential binding sites. However, its original workflow is limited in throughput, sample compatibility and accessible protein targets. Here, we introduce a high-throughput adaptation—HT-PELSA—that increases sample processing efficiency by 100-fold while maintaining high sensitivity and reproducibility. HT-PELSA substantially extends the capabilities of the original method by enabling sensitive protein-ligand profiling in crude cell, tissue and bacterial lysates, allowing the identification of membrane protein targets in diverse biological systems. We demonstrate that HT-PELSA can precisely and accurately determine binding affinities of small molecule inhibitors, sensitively detect direct and allosteric ATP binding sites, and reveal off-target interactions of a marketed kinase inhibitor in heart tissue. By enhancing scalability, reducing costs and enabling system-wide drug screening across a wide range of sample types, HT-PELSA—when combined with next-generation mass spectrometry—may offer a powerful platform poised to accelerate both drug discovery and basic biological research. |
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| Item Description: | Online veröffentlicht: 5. November 2025 Gesehen am 09.02.2026 |
| Physical Description: | Online Resource |
| ISSN: | 1545-9985 |
| DOI: | 10.1038/s41594-025-01699-y |