Induced ubiquitination of the partially disordered estrogen receptor alpha via a 14-3-3 directed molecular glue-PROTAC.

Verhoef, Carlo J A; Crowe, Charlotte; Nakasone, Mark A; DeLaCuadra-Basté, Aitana; Harzing, Tessa; Span, Naomi A S; Sathe, Gajanan; Demmers, Laura C et al. · Nat Commun · 2026

basic_science · Level V

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Abstract

Proteins lacking defined ligandable pockets remain challenging drug targets. Here, we develop a molecular glue-based PROTAC (<sup>MG</sup>PROTAC) approach that chemically conjugates a molecular glue stabilizer to a VHL-recruiting ligand to capture and ubiquitinate the 14-3-3/Estrogen receptor α (ERα) complex. Our designed <sup>MG</sup>PROTACs engage a composite interface between 14-3-3 and the disordered F-domain of ERα, promoting cooperative complex formation and targeted ubiquitination. Biophysical characterization revealed distinct linker-dependent cooperativities across the <sup>MG</sup>PROTAC series, which influenced both cellular permeability and ubiquitination efficiency. Cryo-EM of the most cooperative <sup>MG</sup>PROTAC uncovered de novo VHL-14-3-3ζ contacts, while molecular dynamics simulations rationalize the stabilizing interactions underlying cooperativity. Strikingly, fine-tuning linker design enables selective ubiquitination of distinct complex subunits. These findings establish a structural and mechanistic framework for integrating molecular glue and PROTAC principles, expanding the scope of drug discovery to previously intractable protein complexes.