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

Autor:	Lin H; The Verna and Marrs McLean Department of Biochemistry and Molecular Pharmacology, Baylor College of Medicine, Houston, Texas 77030, United States.; Center for NextGen Therapeutics, Baylor College of Medicine, Houston, Texas 77030, United States., Riching K; Promega Corporation, 2800 Woods Hollow Road, Madison, Wisconsin 53711, United States., Lai MP; Malvern Panalytical Inc., 2400 Computer Drive, Westborough, Massachusetts 01581, United States., Lu D; The Verna and Marrs McLean Department of Biochemistry and Molecular Pharmacology, Baylor College of Medicine, Houston, Texas 77030, United States., Cheng R; The Verna and Marrs McLean Department of Biochemistry and Molecular Pharmacology, Baylor College of Medicine, Houston, Texas 77030, United States.; Center for NextGen Therapeutics, Baylor College of Medicine, Houston, Texas 77030, United States., Qi X; The Verna and Marrs McLean Department of Biochemistry and Molecular Pharmacology, Baylor College of Medicine, Houston, Texas 77030, United States.; Center for NextGen Therapeutics, Baylor College of Medicine, Houston, Texas 77030, United States., Wang J; The Verna and Marrs McLean Department of Biochemistry and Molecular Pharmacology, Baylor College of Medicine, Houston, Texas 77030, United States.; Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas 77030, United States.; Center for NextGen Therapeutics, Baylor College of Medicine, Houston, Texas 77030, United States.
Jazyk:	angličtina
Zdroj:	ACS medicinal chemistry letters [ACS Med Chem Lett] 2024 Jul 28; Vol. 15 (8), pp. 1367-1375. Date of Electronic Publication: 2024 Jul 28 (Print Publication: 2024).
DOI:	10.1021/acsmedchemlett.4c00271
Abstrakt:	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. Competing Interests: The authors declare the following competing financial interest(s): J.W. is a co-founder of Chemical Biology Probes, LLC. and serves as a consultant for CoRegen Inc. (© 2024 American Chemical Society.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu