Maowei Hu, Jason Ochoada, Marisa Actis, Kevin McGowan, Jamie Jarusiewicz, Satoshi Yoshimura, Logan McGrath, Uma Neelakantan, Anup Aggarwal, Anand Mayasundari, Sarah M. Young, Meng Zhang, Lei Yang, Yong Li, Shea Mercer, M. Madan Babu, Marcus Fischer, Brandon Young, Jun Yang, Gisele Nishiguchi, Anang A. Shelat, Daniel J. Blair
High Resolution Image Download MS PowerPoint Slide Molecular glues powerfully control protein proximity but have largely eluded direct screening. A promising avenue for addressing this challenge lies within pinpointing the fundamental features for function-first identification of molecular gluing events. In the widely accepted mechanism, a molecular glue stabilizes two proteins within a ternary complex─here, we show how differences in affinity for ternary and binary complexes directly categorize glues from nonglues. We leverage these differences together with high-throughput chemical synthesis and affinity-selection mass-spectrometry to discover a molecular glue from a suite of over 20,000 crude chemical reaction mixtures. Orthogonal assays robustly support the identification of molecular glues via ternary complex stability. Our findings suggest that a roadmap for de novo molecular glue discovery lies within kinetic profiling of unpurified mixtures of small organic molecules against protein pairs.