PXD025012-2

PXD025012 is an original dataset announced via ProteomeXchange.

Title	‘Off the shelf’ proximity biotinylation for interaction proteomics
Description	Proximity biotinylation workflows typically require CRISPR-based genetic manipulation of target cells. To overcome this bottleneck, we fused the TurboID proximity biotinylation enzyme to Protein A. Upon target cell permeabilization, the ProtA-Turbo enzyme can be targeted to proteins of interest using an endogenous antibody against the bait protein. Addition of biotin then triggers bait-proximal protein biotinylation. Biotinylated proteins can subsequently be enriched from crude lysates and identified by mass spectrometry. We demonstrate this workflow by targeting Emerin, the histone modification H3K9me3 and the chromatin remodeler BRG1. Amongst the main findings, our experiments revealed Flywch1 as an essential protein that interacts with H3K9me3-marked centromeric heterochromatin. The ProtA-Turbo enzyme represents an ‘off the shelf’ proximity biotinylation enzyme that facilitates proximity biotinylation experiments in primary cells and can be used to understand how proteins cooperate in vivo and how this contributes to cellular homeostasis and disease.
HostingRepository	PRIDE
AnnounceDate	2021-07-12
AnnouncementXML	Submission_2021-08-26_06:14:32.153.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Irene Santos Barriopedro
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090; scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive HF; Orbitrap Exploris 480; Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2021-03-25 10:13:55	ID requested
1	2021-07-11 23:01:18	announced
⏵ 2	2021-08-26 06:14:32	announced	2021-08-26: Updated publication reference for PubMed record(s): 34408139.

Santos-Barriopedro I, van Mierlo G, Vermeulen M, Off-the-shelf proximity biotinylation for interaction proteomics. Nat Commun, 12(1):5015(2021) [pubmed]

submitter keyword: Proximity labelling, turboID, protein A, interactome, FLYWCH1

Professor Michiel Vermeulen
contact affiliation	Department of Molecular Biology, Faculty of Science Radboud Institute for Molecular Like Sciences, Radboud University Nijmegen, Nijmegen, 6525 GA, The Netherlands (lab head)
contact email	Michiel.Vermeulen@science.ru.nl
lab head
Irene Santos Barriopedro
contact affiliation	Radboud Universiteit
contact email	I.Santos@science.ru.nl
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/2021/07/PXD025012

PRIDE project URI

• PRIDE
  1. PXD025012
     1. Label: PRIDE project
     2. Name: ‘Off the shelf’ proximity biotinylation for interaction proteomics