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Discovery and optimisation of a covalent ligand for TRIM25 and its application to targeted protein ubiquitination

ABSTRACT: The tripartite motif (TRIM) family of RING-type E3 ligases catalyses the formation of many different types of ubiquitin chains, and as such, plays important roles in diverse cellular functions, ranging from immune regulation to cancer signalling pathways. Few ligands have been discovered for TRIM E3 ligases, and these E3s are under-represented in the rapidly expanding field of induced proximity. Here we present the identification of a novel covalent ligand for the PRYSPRY substrate binding domain of TRIM25. We employ covalent fragment screening coupled with high-throughput chemistry direct-to-biology optimisation to efficiently elaborate covalent fragment hits. We demonstrate that our optimised ligand enhances the in vitro auto-ubiquitination activity of TRIM25 and engages TRIM25 in live cells. We also present the X-ray crystal structure of TRIM25 PRYSPRY in complex with this covalent ligand. Finally, we incorporate our optimised ligand into heterobifunctional proximity-inducing compounds and demonstrate the in vitro targeted ubiquitination of a neosubstrate by TRIM25

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Monocyte, Cell Culture

SUBMITTER: Katherine McPhie

LAB HEAD: Katrin Rittinger

PROVIDER: PXD061182 | Pride | 2025-05-19

REPOSITORIES: Pride

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			Action	DRS
	04113_F1_R1_P0396036B01_TMTPRO16.mgf.bz2	Mgf
	04113_F1_R1_P0396036B01_TMTPRO16.raw.rar	Raw
	04113_F1_R1_P0396036B02_TMTPRO16.mgf.bz2	Mgf
	04113_F1_R1_P0396036B02_TMTPRO16.raw.rar	Raw
	04113_F1_R1_P0396036B03_TMTPRO16.mgf.bz2	Mgf

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