PXD066563-1

PXD066563 is an original dataset announced via ProteomeXchange.

Title	Selective ubiquitination of drug-like small molecules by the ubiquitin ligase HUWE1
Description	The ubiquitin system regulates eukaryotic physiology by modifying specific substrate proteins. Substrate recruitment and the assembly of ubiquitin signals are determined by ubiquitin ligases, some of which also modify non-protein biomolecules. Here we expand this substrate realm, revealing that the human ubiquitin ligase HUWE1 can target drug-like small molecules. We demonstrate that compounds previously reported as HUWE1 inhibitors present substrates of their target ligase. Compound ubiquitination is driven by the canonical catalytic cascade, linking ubiquitin to the compound’s primary amino group. In vitro, the modification is selectively catalyzed by HUWE1, allowing the compounds to compete with protein substrates. We establish cellular detection methods, confirming HUWE1 promotes – but does not exclusively drive – compound ubiquitination in cells. Converting the existing compounds into specific HUWE1 substrates or inhibitors thus requires enhanced specificity. More broadly, our findings open avenues for harnessing the ubiquitin system to convert exogenous small molecules into novel chemical modalities within cells.
HostingRepository	PRIDE
AnnounceDate	2025-09-02
AnnouncementXML	Submission_2025-09-02_12:27:19.640.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Wieland Steinchen
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Synapt MS

Revision	Datetime	Status	ChangeLog Entry
0	2025-07-24 13:04:34	ID requested
⏵ 1	2025-09-02 12:27:20	announced

Dataset with its publication pending

ProteomeXchange project tag: Hydrogen Deuterium Exchange (HDX-MS)

submitter keyword: small-molecule inhibitor,HECT E3, mass spectrometry, enzymology, click chemistry, non-protein ubiquitination

Wieland Steinchen
contact affiliation	Center for Synthetic Microbiology & Department of Chemistry, Marburg University,Hans-Meerwein-Straße 4, 35043 Marburg, Germany
contact email	wieland.steinchen@synmikro.uni-marburg.de
lab head
Wieland Steinchen
contact affiliation	Philipps-University Marburg, Department of Chemistry & SYNMIKRO, Karl-von-Frisch-Straße 14, 35043 Marburg, GERMANY
contact email	wieland.steinchen@synmikro.uni-marburg.de
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2025/09/PXD066563

PRIDE project URI

• PRIDE
  1. PXD066563
     1. Label: PRIDE project
     2. Name: Selective ubiquitination of drug-like small molecules by the ubiquitin ligase HUWE1