PXD055649-1

PXD055649 is an original dataset announced via ProteomeXchange.

Title	Proteomic Profiling of Potential E6AP Substrates via Ubiquitin-based Photo-Crosslinking Assisted Affinity Enrichment
Description	E6AP/UBE3A, the founding member of HECT-type (Homologous to E6AP C-terminus) E3 ligases, is implicated in human papillomaviruses (HPV)-mediated cervical cancer and neurodevelopmental disorders like Angelman and Dup15q syndromes. To elucidate the function of E3 ligases, knowledge about their substrates is crucial. Their identification, however, is challenging due to complex cross-reactivities between E2, E3 and their substrates. To address this, we developed a novel ubiquitin-based photo labeling approach for HECT-type ligases. Our method modifies ubiquitin with a diazirine moiety and attaches it to E6AP's active site using the ubiquitination cascade and a Cys to Lys mutation. The resulting E6AP-Ub conjugate enables substrate trapping via photoaffinity labeling and proteomic profiling. We validated this approach through literature comparison and identified Ub C-terminal hydrolase 7 (Uchl5/Uch37) as a new, proteasome-independent ubiquitination target of E6AP. This method expands the toolbox for substrate identification, potentially providing deeper insights into E6AP-related cellular processes and other HECT-type E3 ligases.
HostingRepository	PRIDE
AnnounceDate	2025-01-27
AnnouncementXML	Submission_2025-01-27_07:36:01.737.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Julian Schuck
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	acetylated residue; monohydroxylated residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2024-09-07 00:17:48	ID requested
⏵ 1	2025-01-27 07:36:02	announced

10.1002/CBIC.202400831;

submitter keyword: photoaffinity labelling, proteomics, substrate identification, ubiquitination,E6AP/UBE3A

Andreas Marx
contact affiliation	University of Constance, Department of Chemistry
contact email	andreas.marx@uni-konstanz.de
lab head
Julian Schuck
contact affiliation	Univerity of Constance
contact email	julian.schuck@uni-konstanz.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/2025/01/PXD055649

PRIDE project URI

• PRIDE
  1. PXD055649
     1. Label: PRIDE project
     2. Name: Proteomic Profiling of Potential E6AP Substrates via Ubiquitin-based Photo-Crosslinking Assisted Affinity Enrichment