PXD061182

PXD061182 is an original dataset announced via ProteomeXchange.

Title	Discovery and optimisation of a covalent ligand for TRIM25 and its application to targeted protein ubiquitination
Description	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
HostingRepository	PRIDE
AnnounceDate	2025-05-19
AnnouncementXML	Submission_2025-05-19_10:49:07.045.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Katherine McPhie
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Eclipse

Revision	Datetime	Status	ChangeLog Entry
0	2025-02-25 08:06:42	ID requested
⏵ 1	2025-05-19 10:49:07	announced

10.1039/D5SC01540E;

submitter keyword: covalent ligand,E3 Ligase, ubiquitin, IA-DTB

Katrin Rittinger
contact affiliation	Molecular Structure of Cell Signalling Laboratory, The Francis Crick Institute
contact email	katrin.rittinger@crick.ac.uk
lab head
Katherine McPhie
contact affiliation	The Francis Crick Institute
contact email	katherine.mcphie@crick.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD061182
     1. Label: PRIDE project
     2. Name: Discovery and optimisation of a covalent ligand for TRIM25 and its application to targeted protein ubiquitination