Target protein localization and its impact on PROTAC-mediated degradation

Simpson, Luke; Glennie, Lorraine; Brewer, Abigail; Zhao, Jin‐Feng; Crooks, Jennifer; Shpiro, Natalia; Sapkota, Gopal P.

doi:10.1016/j.chembiol.2022.08.004

Cited by 45 publications

(31 citation statements)

References 108 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The modularity of the system can be explored further by the addition of protein localization tags to dTAG-SpyCatcher to limit its expression to selected cellular compartments and hence allow for selective depletion of specific pools of the target protein. An analogous approach recently demonstrated the degradability of localization-tagged dTAG in the nucleus, cytoplasm, Golgi, ER, peroxisome, lysosome, and outer mitochondrial membrane . Localization tags on dTAG-SpyCatcher, when matched with the localization of the target protein expression, could also further improve the labeling efficiency and target degradation.…”

Section: Discussionmentioning

confidence: 99%

“…An analogous approach recently demonstrated the degradability of localization-tagged dTAG in the nucleus, cytoplasm, Golgi, ER, peroxisome, lysosome, and outer mitochondrial membrane. 23 Localization tags on dTAG-SpyCatcher, when matched with the localization of the target protein expression, could also further improve the labeling efficiency and target degradation. This versatility makes HiBiT-SpyTag a valuable tool for studying protein biology and the functional consequences of protein degradation in living cells.…”

Section: ■ Discussionmentioning

confidence: 99%

“…While targeted protein degradation has not been previously demonstrated for IRE1α, proteins restricted to the ER have been successfully degraded using dTAG or HaloPROTAC approaches. 23 IRE1α is involved in the ER stress response, and has been studied as a potential anticancer target. 24 Possessing both kinase and RNase domains, IRE1α has been subject to extensive small molecule inhibitor drug discovery efforts against both activities; however, with limited success, possibly due to additional scaffolding or sensory functions not addressed by these approaches.…”

Section: ■ Introductionmentioning

confidence: 99%

“…Additionally, we used SpyTag-HiBiT to tag the C-terminus of IRE1α, an endoplasmic reticulum (ER) membrane protein. While targeted protein degradation has not been previously demonstrated for IRE1α, proteins restricted to the ER have been successfully degraded using dTAG or HaloPROTAC approaches . IRE1α is involved in the ER stress response, and has been studied as a potential anticancer target .…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

HiBiT-SpyTag: A Minimal Tag for Covalent Protein Capture and Degrader Development

et al. 2023

View full text Add to dashboard Cite

The ability to rapidly and selectively modulate cellular protein levels using small molecules is essential for studying complex biological systems. Degradation tags, such as dTAG, allow for selective protein removal with a specific degrader molecule, but their utility is limited by the large tag size (>12 kDa) and the low efficiency of fusion product gene knock-in. Here, we describe the development of a short 24 amino acid peptide tag that enables cellbased quantification and covalent functionalization of proteins to which it is fused. The minimalistic peptide, termed HiBiT-SpyTag, incorporates the HiBiT peptide for protein level quantification and SpyTag, which forms a spontaneous isopeptide bond in the presence of the SpyCatcher protein. Transient expression of dTAG-SpyCatcher efficiently labels HiBiT-SpyTag-modified BRD4 or IRE1α in cells, and subsequent treatment with the dTAG13 degrader results in efficient protein removal without the need for full dTAG knock-in. We also demonstrate the utility of HiBiT-SpyTag for validating the degradation of the endoplasmic reticulum (ER) stress sensor IRE1α, which led to the development of the first PROTAC degrader of the protein. Our modular HiBiT-SpyTag system represents a valuable tool for the efficient development of degraders and for studying other proximity-induced pharmacology.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: ■ Discussionmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

HiBiT-SpyTag: A Minimal Tag for Covalent Protein Capture and Degrader Development

et al. 2023

View full text Add to dashboard Cite

show abstract

“…Notably, the intracellular location of POI should also be taken into account to select the optimal E3 ligase recruiter. It was demonstrated that the alteration of the E3 ligase recruiter depending on the subcellular region where POIs are located can enlarge the degrading efficiency of PROTACs [ 28 ]. This is mainly attributed to each type of E3 ligase being differently distributed in subcellular regions.…”

Section: Protac For Anticancer Therapymentioning

confidence: 99%

Cancer-Specific Delivery of Proteolysis-Targeting Chimeras (PROTACs) and Their Application to Cancer Immunotherapy

et al. 2023

View full text Add to dashboard Cite

Proteolysis-targeting chimeras (PROTACs) are rapidly emerging as a potential therapeutic strategy for cancer therapy by inducing the degradation of tumor-overexpressing oncogenic proteins. They can specifically catalyze the degradation of target oncogenic proteins by recruiting E3 ligases and utilizing the ubiquitin-proteasome pathway. Since their mode of action is universal, irreversible, recyclable, long-lasting, and applicable to ‘undruggable’ proteins, PROTACs are gradually replacing the role of conventional small molecular inhibitors. Moreover, their application areas are being expanded to cancer immunotherapy as various types of oncogenic proteins that are involved in immunosuppressive tumor microenvironments. However, poor water solubility and low cell permeability considerably restrict the pharmacokinetic (PK) property, which necessitates the use of appropriate delivery systems for cancer immunotherapy. In this review, the general characteristics, developmental status, and PK of PROTACs are first briefly covered. Next, recent studies on the application of various types of passive or active targeting delivery systems for PROTACs are introduced, and their effects on the PK and tumor-targeting ability of PROTACs are described. Finally, recent drug delivery systems of PROTACs for cancer immunotherapy are summarized. The adoption of an adequate delivery system for PROTAC is expected to accelerate the clinical translation of PROTACs, as well as improve its efficacy for cancer therapy.

show abstract

Targeting RNA‐binding proteins with small molecules: Perspectives, pitfalls and bifunctional molecules

Kang

2023

FEBS Letters

View full text Add to dashboard Cite

RNA‐binding proteins (RBPs) play vital roles in organisms through binding with RNAs to regulate their functions. Small molecules affecting the function of RBPs have been developed, providing new avenues for drug discovery. Herein, we describe the perspectives on developing small molecule regulators of RBPs. The following types of small molecule modulators are of great interest in drug discovery: small molecules binding to RBPs to affect interactions with RNA molecules, bifunctional molecules binding to RNA or RBP to influence their interactions, and other types of molecules that affect the stability of RNA or RBPs. Moreover, we emphasize that the bifunctional molecules may play important roles in small molecule development to overcome the challenges encountered in the process of drug discovery.

show abstract

Target protein localization and its impact on PROTAC-mediated degradation

Cited by 45 publications

References 108 publications

HiBiT-SpyTag: A Minimal Tag for Covalent Protein Capture and Degrader Development

HiBiT-SpyTag: A Minimal Tag for Covalent Protein Capture and Degrader Development

Cancer-Specific Delivery of Proteolysis-Targeting Chimeras (PROTACs) and Their Application to Cancer Immunotherapy

Targeting RNA‐binding proteins with small molecules: Perspectives, pitfalls and bifunctional molecules

Contact Info

Product

Resources

About