PXD005758-1

PXD005758 is an original dataset announced via ProteomeXchange.

Title	A method for spatially and temporally resolved protein network interrogation in living cells
Description	Cells operate through protein interaction networks organized in space and in time, but current methods to interrogate such biology typically resolve only one of these dimensions. Here, we describe a method to resolve both dimensions simultaneously using proximity labeling mediated by engineered ascorbic acid peroxidase (APEX). APEX has been used to label organelle proteomes with high temporal resolution, but its spatial resolution is generally thought to be limited because of its larger labeling radius. We describe an analytical pipeline, based on quantitative proteomics using spatial references, which overcomes this barrier. As proof-of-principle, we apply the methodology to interrogate protein interaction networks of G protein coupled receptors both temporally and spatially. Using this approach, we identify known binding partners as well as novel receptor regulators. Validating the methodology as a discovery tool, we demonstrate that two of these components drive ubiquitin-linked down-regulation of opioid receptors.
HostingRepository	PRIDE
AnnounceDate	2017-03-03
AnnouncementXML	Submission_2017-03-03_02:04:42.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Ruth Huttenhain
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion; LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2017-01-21 07:53:50	ID requested
⏵ 1	2017-03-03 02:04:43	announced
2	2024-10-22 04:32:20	announced	2024-10-22: Updated project metadata.

Dataset with its publication pending

curator keyword: Technical, Biological

submitter keyword: GPCR, APEX, proximity biotinylation, protein-protein interactions

Ruth Huttenhain
contact affiliation	Department of Molecular & Cellular Pharmacology UCSF, San Francisco, CA
contact email	ruth.huttenhain@ucsf.edu
lab head
Ruth Huttenhain
contact affiliation	UCSF
contact email	ruth.huttenhain@ucsf.edu
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/2017/03/PXD005758

PRIDE project URI

• PRIDE
  1. PXD005758
     1. Label: PRIDE project
     2. Name: A method for spatially and temporally resolved protein network interrogation in living cells