PXD074567

PXD074567 is an original dataset announced via ProteomeXchange.

Title	BromoCatch: a self-labelling tag platform for protein analysis and live cell imaging
Description	Visualizing and manipulating proteins in live cells is crucial for studying complex biological processes. Self-labelling protein (SLP) tags such as HaloTag and SNAP-tag can be fused to genes of interest to allow protein labelling in cells. New tools to enable rapid, specific and stable protein labelling are in demand to expand the arsenal of SLPs. Here we present BromoCatch, a novel SLP platform based on a small ~13 kDa bromodomain (BD) engineered with a nucleophilic cysteine for covalent ligand engagement. A structure-based designed library of 16 “bumped” binders bearing diverse electrophilic warheads was screened against two cysteine mutants using differential scanning fluorimetry and intact protein mass spectrometry to monitor covalent complex formation. We qualify the para-acrylamide bumped derivative MR116 and the Brd4-BD2 double mutant L387A,E438C as the protein-ligand pair of choice, and its covalent binding mode was confirmed by an X-ray cocrystal structure solved to 1.3 Å of resolution. The BromoCatch platform exhibited potent and irreversible target engagement in cells using nanoBRET and residence time assays. Practicality and scope are further demonstrated through the design and proof-of-concept application of a biotinylated conjugate, PROTAC tag degraders, and fluorescent probes of both full-on and switch-on types for ex-cellulo and live-cell imaging, including side-by-side comparison and orthogonality with HaloTag. Together, we qualify BromoCatch as a novel, versatile and efficient protein labelling tool. Its advantageous design features and kinetic fitness enable modular attachment of diverse functionalities, multiplexing with other SLPs, and catalytic degradation, which will open future applications and witness broad utility.
HostingRepository	PRIDE
AnnounceDate	2026-05-04
AnnouncementXML	Submission_2026-05-04_07:10:37.621.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Gajanan Sathe
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Ascend

Revision	Datetime	Status	ChangeLog Entry
0	2026-02-17 16:02:08	ID requested
⏵ 1	2026-05-04 07:10:38	announced

Dataset with its publication pending

submitter keyword: BromoCatch, cell imaging

Alessio Ciulli
contact affiliation	Professor Centre for Targeted Protein Degradation, School of Life Sciences, university of Dundee
contact email	a.ciulli@dundee.ac.uk
lab head
Gajanan Sathe
contact affiliation	Centre for Targeted Protein Degradation (CeTPD) University of Dundee
contact email	gsathe001@dundee.ac.uk
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/2026/05/PXD074567

PRIDE project URI

• PRIDE
  1. PXD074567
     1. Label: PRIDE project
     2. Name: BromoCatch: a self-labelling tag platform for protein analysis and live cell imaging