Biology subjects

Gui, W.

Publications and source records attributed to Gui, W..

3 recordsLinked to original sources

bioRxiv2026

A Proximity-Driven Functional Screening Platform for the Discovery of Noninhibitory USP7 Ligand Enabling Targeted Protein Stabilization

Targeted protein stabilization by deubiquitinase-targeting chimeras (DUBTACs) has emerged as a promising therapeutic strategy for preserving the function of essential proteins. DUBTACs are bifunctional molecules that recruit deubiquitinating enzymes (DUBs) to remove ubiquitin chains from substrate proteins, thereby preventing their proteasomal degradation. A key requirement for DUBTACs development is the identification of ligands that bind DUBs without impairing their catalytic activity. However, such noninhibitory ligands are exceedingly rare. Here, we report a novel proximity-driven on-bead functional screening platform for the discovery of noninhibitory DUB ligands. Using this approach, we discovered a previously unreported noninhibitory ligand for USP7. Furthermore, we generated DUBTACs by conjugating this USP7 ligand to a CFTR-binding ligand. Our lead compound, WJ045, effectively stabilized CFTR protein levels in cells, demonstrating the potential of this platform for discovering DUB recruiters and facilitating the development of targeted protein stabilization therapeutics.

biochemistry↗

bioRxiv2025

A Functional Assay For Mining Non-Inhibitory Enzyme Ligands From One Bead One Compound Libraries: Application to E3 Ubiquitin Ligases

Chemical dimerizers are synthetic molecules that bring into proximity two or more proteins that do not normally interact with one another. A major application of this technology is to recruit an enzyme to a target protein, resulting in its post-translational modification (PTM). In particular, chemical dimerizer-mediated poly-Ubiquitylation of proteins has garnered an enormous amount of interest as a new drug modality. A fundamental requirement for the construction of new PTM-driving dimerizers is an enzyme ligand that does not inhibit its activity. Traditional activity-based high-throughput screening platforms are not suited for this purpose. Here we describe a novel platform for screening libraries of bead-displayed compounds that links a requirement for small molecule binding to the enzyme with enzyme-mediated modification of a nearby substrate. This system ensures that the enzyme-recruiting small molecules do not interfere with the catalytic function of the enzyme. We demonstrate the utility of this system in the context of E3 Ubiquitin ligase-recruiting molecules and report the discovery of a novel, low molecular mass ligand for the Von Hippel Landau (VHL) protein.

biochemistry↗

bioRxiv2023

Chemically Induced Degradation of Native Proteins by Direct Recruitment to the 26S Proteasome

Targeted protein degradation (TPD) is a promising strategy for drug development. Most degraders function by forcing the association of the target protein (TP) with an E3 Ubiquitin ligase, which in favorable cases results in the poly-Ubiquitylation of the TP and its subsequent degradation by the 26S proteasome. Here we explore the feasibility of a different TPD strategy in which the TP is recruited directly to the proteasome without the requirement for poly-Ubiquitylation. Using an engineered cell line in which the HaloTag protein is fused to one of the Ubiquitin receptors, we show that native protein targets can be degraded in this fashion when the cells are exposed to a chemical dimerizer containing a chloroalkane and a TP ligand. The potential advantages and disadvantages of Ubiquitin-independent degraders vs. traditional proteolysis-targeting chimeras are discussed.

molecular biology↗