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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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Hauptverfasser: Li, Kejia (VerfasserIn) , Potel, Clément M. (VerfasserIn) , Becher, Isabelle (VerfasserIn) , Hüttmann, Nico (VerfasserIn) , Garrido-Rodriguez, Martin (VerfasserIn) , Schwarz, Jennifer Jasmin (VerfasserIn) , Burtscher, Mira Lea (VerfasserIn) , Savitski, Mikhail M. (VerfasserIn)
Dokumenttyp: Article (Journal)
Sprache:Englisch
Veröffentlicht: 2026
In: Nature structural & molecular biology
Year: 2026, Jahrgang: 33, Heft: 1, Pages: 184-192
ISSN:1545-9985
DOI:10.1038/s41594-025-01699-y
Online-Zugang: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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Verfasserangaben:Kejia Li, Clement M. Potel, Isabelle Becher, Nico Hüttmann, Martin Garrido-Rodriguez, Jennifer Schwarz, Mira Lea Burtscher, Mikhail M. Savitski
Beschreibung
Zusammenfassung: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.
Beschreibung:Online veröffentlicht: 5. November 2025
Gesehen am 09.02.2026
Beschreibung:Online Resource
ISSN:1545-9985
DOI:10.1038/s41594-025-01699-y

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