PXD068381

PXD068381 is an original dataset announced via ProteomeXchange.

Title	Proximity labelling identification of ULK1 wild-type (WT) and ULK1-K46I (kinase dead) interactors upon mitochondria tethering in U2OS WT cells
Description	This project involved generating three stable cell lines using a Flp-In system, each containing specific integrated constructs: "FKBPx2-myc-APEX2-ULK1", "FKBPx2-myc-APEX2-ULK1-K46I" (kinase-dead mutant), and "FKBPx2-myc-APEX2" (control). All cell lines additionally express an outer mitochondrial membrane-targeted construct consisting of truncated FIS193–152 fused to FRB (FRB-FIS193–152). Upon treatment with a heterodimerizer (rapalog), the cytosolic FKBP-tagged constructs are recruited to mitochondria via FRB-rapalog-FKBP interactions, enabling controlled mitochondrial tethering. The constructs contain APEX2 peroxidase, which catalyzes biotin-phenol biotinylation of nearby proteins upon H₂O₂ treatment, allowing proximity-dependent protein identification. The primary objective was to investigate the interactomes of wild-type ULK1 versus the kinase-dead ULK1-K46I mutant when localized to mitochondria. By comparing the biotinylated protein profiles between these conditions and the APEX2-only control, we aimed to identify ULK1-specific interacting partners and determine how kinase activity affects these interactions in the mitochondrial environment. This approach provides insights into ULK1's role in mitophagy and autophagy regulation.
HostingRepository	PRIDE
AnnounceDate	2026-04-24
AnnouncementXML	Submission_2026-04-24_04:36:00.818.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	David Hollenstein
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606;
ModificationList	iodoacetamide derivatized residue
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2025-09-15 06:53:09	ID requested
⏵ 1	2026-04-24 04:36:01	announced

10.1242/JCS.264577;

submitter keyword: mitophagy, autophagy, ULK1-K46I, ULK1, proximity labeling,APEX2

Prof. Dr. Claudine Kraft
contact affiliation	Institute of Biochemistry and Molecular Biology Department of Biochemistry, University of Freiburg, Germany
contact email	claudine.kraft@biochemie.uni-freiburg.de
lab head
David Hollenstein
contact affiliation	Mass Spectrometry Facility, Max Perutz Laboratories, Vienna Biocenter Campus (VBC), Dr. Bohr-Gasse 7, Vienna 1030, Austria
contact email	hollenstein.david@gmail.com
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/2026/04/PXD068381

PRIDE project URI

• PRIDE
  1. PXD068381
     1. Label: PRIDE project
     2. Name: Proximity labelling identification of ULK1 wild-type (WT) and ULK1-K46I (kinase dead) interactors upon mitochondria tethering in U2OS WT cells