PXD066625

PXD066625 is an original dataset announced via ProteomeXchange.

Title	ASAP-ID: Proximity labelling with small tags
Description	Biotinylation-based proximity labelling methods are valuable for discovering protein-protein interactions within cellular systems. However, one limitation of these approaches is that most require fusing the target protein with the enzyme that biotinylates nearby proteins (i.e., TurboID or APEX2), which risks sterically disrupting the protein's native function. Here, we present a method designed to reduce the steric impact of these fusions and offer greater flexibility in labelling modalities. The method, Antibody and Small-tag Assembly on Proteins for Interaction Detection (ASAP-ID), involves a bipartite system. Target proteins are fused to a small peptide antigen that recruits TurboID or APEX2 fused to an antibody directed to the antigen. Using two different antigen/antibody systems (SunTag and MoonTag), we show that ASAP-ID can specifically label human Lamin A in cells. We also demonstrate that the approach works when the antibody fusion is added in-trans to fixed cells post-expression (ASAP-IDIT). ASAP-IDIT identified more than 448 known and previously undescribed interactors of lamin. We further used ASAP-IDIT to study how ALS-mutant profilin 1 affected its interactome. The method identified proteins involved in protein quality control that correlated with aggregation propensity. Moreover, the different mutants showed variation in the cellular location where aggregates formed. ASAP-IDIT revealed preferences for mitochondrial proteins for the two profilin mutants that tend to aggregate in the cytoplasm, C71G and M114T, and nuclear proteins for a mutant more prone to nuclear aggregation. These findings position ASAP-ID as a powerful addition to the proximity labelling toolkit, capable of probing subtle differences in interactomes in a less invasive manner.
HostingRepository	PRIDE
AnnounceDate	2026-09-07
AnnouncementXML	Submission_2026-09-06_16:32:51.267.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Ruohua Lyu
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606;
ModificationList	acetylated residue; monohydroxylated residue
Instrument	Orbitrap Ascend

Revision	Datetime	Status	ChangeLog Entry
0	2025-07-27 17:27:13	ID requested
⏵ 1	2026-09-06 16:32:52	announced

Lyu R, Ruff KM, Palmer CS, Ramirez AE, Ormsby AR, Scott DJ, Pappu RV, Stojanovski D, Stroud DA, Hatters DM, ASAP-ID: Proximity Labelling With Small Tags. Mol Cell Proteomics, 25(8):101616(2026) [pubmed]

10.1016/j.mcpro.2026.101616;

submitter keyword: Human, LC-MS/MS

Danny Hatters
contact affiliation	Department of Biochemistry and Pharmacology, Bio21 Molecular Science and Biotechnology Institute. The University of Melbourne.
contact email	dhatters@unimelb.edu.au
lab head
Ruohua Lyu
contact affiliation	The University of Melbourne
contact email	helen.lyu@unimelb.edu.au
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/2026/09/PXD066625

PRIDE project URI

• PRIDE
  1. PXD066625
     1. Label: PRIDE project
     2. Name: ASAP-ID: Proximity labelling with small tags