PXD025775

PXD025775 is an original dataset announced via ProteomeXchange.

Title	Endosomal cAMP production impacts the broad cellular phosphoproteome
Description	Endosomal signaling from G protein-coupled receptors (GPCRs) has emerged as a novel paradigm with important pharmacological and physiological implications. Yet, our knowledge of the functional consequences of activating intracellular GPCRs is incomplete. To address this gap, we combined an optogenetic approach for site-specific generation of the prototypical second messenger cyclic AMP (cAMP) with unbiased phosphoproteomic analysis. We identified 218 unique sites that either increased or decreased in phosphorylation upon cAMP production. We next determined that the compartment of signaling origin impacted the regulation of the entire repertoire of targets in that, remarkably, endosome-derived cAMP led to more robust changes in phosphorylation for all targets regardless of their annotated sub-cellular localization. Furthermore, we observed that proteins that are dephosphorylated in response to cAMP accumulation exhibited disproportionately strong bias towards endosomal over plasma membrane signaling. Through bioinformatics analysis, we established that this specific set of targets are substrates for protein phosphatase 2A, PP2A-B56δ, and propose compartmentalized activation of PP2A as the likely underlying mechanism. Altogether, our study extends the concept that endosomal signaling is a significant functional contributor to cellular responsiveness by establishing a unique role for localized cAMP production in defining categorically distinct phosphoresponses.
HostingRepository	PRIDE
AnnounceDate	2021-06-15
AnnouncementXML	Submission_2021-06-14_23:25:35.796.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Jeffrey Johnson
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	phosphorylated residue
Instrument	LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2021-05-03 23:09:02	ID requested
⏵ 1	2021-06-14 23:25:36	announced

Dataset with its publication pending

submitter keyword: cAMP- cyclic AMP

bPAC- bacterial photoactivatable adenylyl cyclase, GPCR- G protein-coupled receptor

SILAC- stable isotope labeling with amino acids in cell culture

Fe3+-NTA IMAC- iron (III)-nitrilotriacetic acid immobilized metal ion affinity

LC-MS/MS- liquid chromatography-tandem mass spectrometry

PKA- protein kinase A

PDE- phosphodiesterase

PP2A- protein phosphatase 2A

Mark von Zastrow
contact affiliation	University of California San Francisco
contact email	mark@vzlab.org
lab head
Jeffrey Johnson
contact affiliation	Icahn School of Medicine at Mount Sinai
contact email	jeffrey.johnson@mssm.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/2021/06/PXD025775

PRIDE project URI

• PRIDE
  1. PXD025775
     1. Label: PRIDE project
     2. Name: Endosomal cAMP production impacts the broad cellular phosphoproteome