A caged E3 ligase ligand for PROTAC-mediated protein degradation with light

A caging group has been appended to a widely used Von Hippel Lindau (VHL) E3 ligase ligand for targeted protein degradation with PROTACs. Proteolysis is triggered only after a short irradiation time allowing spatiotemporal control of the protein’s fate.

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A Phenotypic Approach for the Identification of New Molecules for Targeted Protein Degradation Applications

SLAS DISCOVERY Advancing Life Sciences ◽

10.1177/24725552211017517 ◽

2021 ◽

pp. 247255522110175

Author(s):

Peter Stacey ◽

Hannah Lithgow ◽

Xiao Lewell ◽

Agnieszka Konopacka ◽

Stephen Besley ◽

...

Keyword(s):

Fusion Protein ◽

Protein Degradation ◽

E3 Ligase ◽

Ubiquitin Proteasome System ◽

E3 Ligases ◽

Von Hippel Lindau ◽

Protein Levels ◽

New Strategy ◽

Cellular Context

Targeted protein degradation is an emerging new strategy for the modulation of intracellular protein levels with applications in chemical biology and drug discovery. One approach to enable this strategy is to redirect the ubiquitin–proteasome system to mark and degrade target proteins of interest (POIs) through the use of proteolysis targeting chimeras (PROTACs). Although great progress has been made in enabling PROTACs as a platform, there are still a limited number of E3 ligases that have been employed for PROTAC design. Herein we report a novel phenotypic screening approach for the identification of E3 ligase binders. The key concept underlying this approach is the high-throughput modification of screening compounds with a chloroalkane moiety to generate HaloPROTACs in situ, which were then evaluated for their ability to degrade a GFP-HaloTag fusion protein in a cellular context. As proof of concept, we demonstrated that we could generate and detect functional HaloPROTACs in situ, using a validated Von Hippel–Lindau (VHL) binder that successfully degraded the GFP-HaloTag fusion protein in living cells. We then used this method to prepare and screen a library of approximately 2000 prospective E3 ligase-recruiting molecules.

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A Caged E3 Ligase Ligand for PROTAC-Mediated Protein Degradation with Light

10.26434/chemrxiv.11350079.v2 ◽

2019 ◽

Author(s):

Cyrille Kounde ◽

Maria M. Shchepinova ◽

Edward Tate

Keyword(s):

Protein Degradation ◽

Irradiation Time ◽

E3 Ligase ◽

Von Hippel Lindau ◽

Short Irradiation ◽

Short Irradiation Time ◽

Spatiotemporal Control

A caging group has been appended to a widely used Von Hippel Lindau (VHL) E3 ligase ligand for targeted protein degradation with PROTACs. Proteolysis is triggered only after a short irradiation time allowing spatiotemporal control of the protein’s fate.

Download Full-text

A Caged E3 Ligase Ligand for PROTAC-Mediated Protein Degradation with Light

10.26434/chemrxiv.11350079.v1 ◽

2019 ◽

Author(s):

Cyrille Kounde ◽

Maria M. Shchepinova ◽

Edward Tate

Keyword(s):

Protein Degradation ◽

Irradiation Time ◽

E3 Ligase ◽

Von Hippel Lindau ◽

Short Irradiation ◽

Short Irradiation Time ◽

Spatiotemporal Control

A caging group has been appended to a widely used Von Hippel Lindau (VHL) E3 ligase ligand for targeted protein degradation with PROTACs. Proteolysis is triggered only after a short irradiation time allowing spatiotemporal control of the protein’s fate.

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PHOTACs Enable Optical Control of Protein Degradation

10.26434/chemrxiv.8206688 ◽

2019 ◽

Cited By ~ 2

Author(s):

Martin Reynders ◽

Bryan Matsuura ◽

Marleen Bérouti ◽

Daniele Simoneschi ◽

Antonio Marzio ◽

...

Keyword(s):

Protein Degradation ◽

E3 Ligase ◽

Optical Control ◽

Cellular Proteins ◽

Bifunctional Molecules ◽

Protein Levels ◽

Lead Compounds ◽

Subsequent Degradation ◽

Temporal And Spatial ◽

Precision Therapeutics

PROTACs (proteolysis targeting chimeras) are bifunctional molecules that tag proteins for ubiquitylation by an E3 ligase complex and subsequent degradation by the proteasome. They have emerged as powerful tools to control the levels of specific cellular proteins and are on the verge of being clinically used. We now introduce photoswitchable PROTACs that can be activated with the temporal and spatial precision that light provides. These trifunctional molecules, which we named PHOTACs, consist of a ligand for an E3 ligase, a photoswitch, and a ligand for a protein of interest. We demonstrate this concept by using PHOTACs that target either BET family proteins (BRD2,3,4) or FKBP12. Our lead compounds display little or no activity in the dark but can be reversibly activated to varying degrees with different wavelengths of light. Our modular and generalizable approach provides a method for the optical control of protein levels with photopharmacology and could lead to new types of precision therapeutics that avoid undesired systemic toxicity.

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Mutated SPOP E3 ligase promotes 17βHSD4 protein degradation to drive androgenesis and prostate cancer progression

Cancer Research ◽

10.1158/0008-5472.can-20-3258 ◽

2021 ◽

pp. canres.3258.2020

Author(s):

Lei Shi ◽

Yuqian Yan ◽

Yundong He ◽

Binyuan Yan ◽

Yunqian Pan ◽

...

Keyword(s):

Prostate Cancer ◽

Protein Degradation ◽

Cancer Progression ◽

E3 Ligase ◽

Prostate Cancer Progression

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A glycine-specific N-degron pathway mediates the quality control of protein N-myristoylation

Science ◽

10.1126/science.aaw4912 ◽

2019 ◽

Vol 365 (6448) ◽

pp. eaaw4912 ◽

Cited By ~ 27

Author(s):

Richard T. Timms ◽

Zhiqian Zhang ◽

David Y. Rhee ◽

J. Wade Harper ◽

Itay Koren ◽

...

Keyword(s):

Quality Control ◽

Protein Stability ◽

Protein Degradation ◽

E3 Ligase ◽

Cleavage Sites ◽

Caspase Cleavage ◽

E3 Ligases ◽

Ring E3 Ligase ◽

The Stability ◽

Ring E3

The N-terminal residue influences protein stability through N-degron pathways. We used stability profiling of the human N-terminome to uncover multiple additional features of N-degron pathways. In addition to uncovering extended specificities of UBR E3 ligases, we characterized two related Cullin-RING E3 ligase complexes, Cul2ZYG11B and Cul2ZER1, that act redundantly to target N-terminal glycine. N-terminal glycine degrons are depleted at native N-termini but strongly enriched at caspase cleavage sites, suggesting roles for the substrate adaptors ZYG11B and ZER1 in protein degradation during apoptosis. Furthermore, ZYG11B and ZER1 were found to participate in the quality control of N-myristoylated proteins, in which N-terminal glycine degrons are conditionally exposed after a failure of N-myristoylation. Thus, an additional N-degron pathway specific for glycine regulates the stability of metazoan proteomes.

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