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Lookup NU author(s): Dr Richard Heath, Professor Wyatt YueORCiD
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY-NC-ND).
© The Author(s) 2025.We report an enantioselective protein affinity selection mass spectrometry screening approach (E-ASMS) that enables the detection of weak binders, informs on selectivity, and generates orthogonal confirmation of binding. After method development with control proteins, we screen 31 human proteins against a designed library of 8,217 chiral compounds. We identify 16 binders to 12 targets, including many proteins predicted to be “challenging to ligand”, and confirm their interactions through orthogonal biophysical assays. Seven binders to six targets display enantioselective binding, with KD values ranging from 3 to 20 µM. Binders for four targets (DDB1, WDR91, WDR55, and HAT1) are selected for in-depth characterization using X-ray crystallography. In all four cases, the mechanisms underlying enantioselectivity are readily explained. These results demonstrate that E-ASMS enables the identification and characterization of selective and weakly binding ligands for novel protein targets with unprecedented throughput and sensitivity.
Author(s): Wang X, Sun J, Ahmad S, Yang D, Li F, Chan UH, Zeng H, Simoben CV, Green SR, Silva M, Houliston S, Dong A, Bolotokova A, Gibson E, Kutera M, Ghiabi P, Kondratov I, Matviyuk T, Chuprina A, Mavridi D, Lenz C, Joerger AC, Brown BD, Heath RB, Yue WW, Robbie LK, Beyett TS, Muller S, Knapp S, Owen DR, Harding R, Schapira M, Brown PJ, Santhakumar V, Ackloo S, Arrowsmith CH, Edwards AM, Peng H, Halabelian L
Publication type: Article
Publication status: Published
Journal: Nature Communications
Year: 2026
Volume: 17
Issue: 1
Online publication date: 17/12/2025
Acceptance date: 26/11/2025
Date deposited: 02/02/2026
ISSN (electronic): 2041-1723
Publisher: Nature Research
URL: https://doi.org/10.1038/s41467-025-67403-2
DOI: 10.1038/s41467-025-67403-2
Data Access Statement: Atomic coordinates and structure factors for all crystal structures have been deposited in the Protein Data Bank under the accession codes: 9EJO, 9EJP, 9EJQ, 9EKP, and 9MJG. All raw data generated in this work have been deposited in the NIH Common Fund’s National Metabolomics Data Repository (NMDR) under Project ID NMDR: PR002550. The dataset is publicly available and can be accessed via the project [https://doi.org/10.21228/M81K02]. Source data are provided with this paper. Code availability The source code and demo datasets can be found via GitHub at [https://github.com/huiUofT/EAS-MS].
PubMed id: 41407727
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