PXD030963-1

PXD030963 is an original dataset announced via ProteomeXchange.

Title	Off the shelf proximity biotinylation using ProtA-TurboID
Description	Proximity biotinylation is a commonly used method to identify the in vivo proximal proteome for proteins of interest. This technology typically relies on fusing a bait protein to a biotin ligase using overexpression or CRISPR-based tagging, thus prohibiting such assays in (primary) cell types that are difficult to transfect. We recently developed an ‘off the shelf’ proximity biotinylation method, which makes use of a recombinant enzyme consisting of the biotin ligase TurboID fused to the antibody-recognizing moiety Protein A. In this method, a bait specific antibody and the ProteinA-Turbo enzyme are consecutively added to permeabilized fixed or unfixed cells. Following incubation, during which ProteinA-Turbo-antibody-antigen complexes are formed, unbound molecules are washed away, after which bait-proximal biotinylation is triggered by the addition of exogenous biotin. Finally, biotinylated proteins are enriched from crude lysates using streptavidin beads followed by mass spectrometry-based protein identification. Here, we present a detailed protocol for this method
HostingRepository	PRIDE
AnnounceDate	2022-10-17
AnnouncementXML	Submission_2022-10-16_23:58:54.881.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	IreneSantos Barriopedro
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2022-01-14 03:52:31	ID requested
⏵ 1	2022-10-16 23:58:55	announced
2	2023-11-14 08:54:48	announced	2023-11-14: Updated project metadata.

submitter keyword: cell lines, centromeres, biotinylation, histone PTMs,Protein-protein interactions

Prof MichielVermeulen
contact affiliation	Department of Molecular Biology, Faculty of Science, Radboud Institute for Molecular Life Sciences, Radboud University Nijmegen, Nijmegen, 6525 GA, The Netherlands (lab head) (lab head)
contact email	michiel.vermeulen@science.ru.nl
lab head
IreneSantos 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/2022/10/PXD030963

PRIDE project URI

• PRIDE
  1. PXD030963
     1. Label: PRIDE project
     2. Name: Off the shelf proximity biotinylation using ProtA-TurboID