Cullin-RING ligases employ geometrically optimized catalytic partners for substrate targeting

Li, Jerry; Purser, Nicholas; Liwocha, Joanna; Scott, Daniel C.; Byers, Holly A.; Steigenberger, Barbara; Hill, Spencer; Tripathi-Giesgen, Ishita; Hinkle, Trent; Hansen, Fynn M.; Prabu, J. Rajan; Radhakrishnan, Senthil K.; Kirkpatrick, Donald S.; Reichermeier, Kurt M.; Schulman, Brenda A.; Kleiger, Gary

doi:10.1016/j.molcel.2024.01.022

Cited by 5 publications

(1 citation statement)

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“…The efficient ubiquitination of a substrate lysine requires the appropriate spatial proximity between the substrate lysine and the narrow catalytic site on the E2 enzyme. − This spatial arrangement is crucial for facilitating the ubiquitination reaction. Therefore, the accessibility and positioning of lysine residues play a critical role in determining their susceptibility to ubiquitination. , It has been reported that lysine residues of the target protein located on the ubiquitin accessible band of the E3 ligase machinery will contribute more to the degradation. − Therefore, recently, there have been increasing concerns that the introduced lysine residues on the tagging protein or peptide may contribute to degradation artifacts.…”

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confidence: 99%

Lysineless HiBiT and NanoLuc Tagging Systems as Alternative Tools for Monitoring Targeted Protein Degradation

Lin,

Riching,

Lai

et al. 2024

ACS Med. Chem. Lett.

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Target protein degradation (TPD) has emerged as a revolutionary approach in drug discovery, leveraging the cell's intrinsic machinery to selectively degrade disease-associated proteins. Nanoluciferase (nLuc) fusion proteins and the NanoBiT technology offer two robust and sensitive screening platforms to monitor the subtle changes in protein abundance induced by TPD molecules. Despite these advantages, concerns have arisen regarding potential degradation artifacts introduced by tagging systems due to the presence of lysine residues on them, prompting the development of alternative tools. In this study, we introduce HiBiT-RR and nLuc K0 , variants devoid of lysine residues, to mitigate such artifacts. Our findings demonstrate that HiBiT-RR maintains a similar sensitivity and binding affinity with the original HiBiT. Moreover, the comparison between nLuc WT and nLuc K0 constructs reveals variations in degradation patterns induced by certain TPD molecules, emphasizing the importance of choosing appropriate tagging systems to ensure the reliability of experimental outcomes in studying protein degradation processes.

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mentioning

confidence: 99%

Lysineless HiBiT and NanoLuc Tagging Systems as Alternative Tools for Monitoring Targeted Protein Degradation

Lin,

Riching,

Lai

et al. 2024

ACS Med. Chem. Lett.

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Diversity of structure and function in Cullin E3 ligases

Lin,

Komives

2024

Current Opinion in Structural Biology

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Extrapolating Lessons from Targeted Protein Degradation to Other Proximity-Inducing Drugs

Winter

2024

ACS Chem. Biol.

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Targeted protein degradation (TPD) is an emerging pharmacologic strategy. It relies on small-molecule “degraders” that induce proximity of a component of an E3 ubiquitin ligase complex and a target protein to induce target ubiquitination and subsequent proteasomal degradation. Essentially, degraders thus expand the function of E3 ligases, allowing them to degrade proteins they would not recognize in the absence of the small molecule. Over the past decade, insights gained from identifying, designing, and characterizing various degraders have significantly enhanced our understanding of TPD mechanisms, precipitating in rational degrader discovery strategies. In this Account, I aim to explore how these insights can be extrapolated to anticipate both opportunities and challenges of utilizing the overarching concept of proximity-inducing pharmacology to manipulate other cellular circuits for the dissection of biological mechanisms and for therapeutic purposes.

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Cullin-RING ligases employ geometrically optimized catalytic partners for substrate targeting

Cited by 5 publications

References 110 publications

Lysineless HiBiT and NanoLuc Tagging Systems as Alternative Tools for Monitoring Targeted Protein Degradation

Lysineless HiBiT and NanoLuc Tagging Systems as Alternative Tools for Monitoring Targeted Protein Degradation

Diversity of structure and function in Cullin E3 ligases

Extrapolating Lessons from Targeted Protein Degradation to Other Proximity-Inducing Drugs

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