PXD027905

PXD027905 is an original dataset announced via ProteomeXchange.

Title	A Network G Protein Gai Signaling Partners is Revealed by Proximity Labeling Proteomics and Includes PDZ-RhoGEF
Description	G protein-coupled receptors (GPCRs) that couple to the Gi family of G proteins are key regulators of cell and tissue physiology. Our previous work has revealed new roles for Gαi in regulating the migration of neutrophils and fibrosarcoma cells downstream of activated chemoattractant receptors. We used an intact cell proximity-based labeling approach using BioID2 coupled to tandem mass tag (TMT)-based quantitative proteomics to identify proteins that selectively interacted with the GTP-bound form of Gαi1. Multiple targets were identified and validated for selective biotinylation by a constitutively active BioID2-tagged Gai1 mutant, suggesting a network of interactions for activated Gαi proteins in intact cells. We showed that active Gαi1 stimulated one candidate protein, PDZ‐RhoGEF (PRG) and that despite over 85% sequence identity, the ability of Gαi1, Gαi2, and Gαi3 isoforms to activate PRG greatly differed. We also demonstrated that active Gαi likely regulates polarized myosin light chain kinase phosphorylation through activation of PRG in primary human neutrophils, suggesting functional relevance of this interaction. Identification and characterization of new targets regulated by Gαi both individually and in networks provide insights that will aid in the investigation of the functional roles of Gi-coupled GPCRs in multiple biological processes.
HostingRepository	PRIDE
AnnounceDate	2022-08-12
AnnouncementXML	Submission_2022-08-12_06:20:30.268.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Naincy Chandan
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	TMT6plex-126 reporter+balance reagent acylated residue; monohydroxylated residue; iodoacetamide derivatized residue; deamidated residue
Instrument	Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2021-08-12 14:43:07	ID requested
⏵ 1	2022-08-12 06:20:31	announced

Chandan NR, Abraham S, SenGupta S, Parent CA, Smrcka AV, signaling partners is revealed by proximity labeling proteomics analysis and includes PDZ-RhoGEF. Sci Signal, 15(717):eabi9869(2022) [pubmed]

submitter keyword: GPCR signaling, Gαi proteins, Proximity labeling, Mass spectrometry, Interactome, PDZ-RhoGEF, Cell-migration

Alan V. Smrcka
contact affiliation	Department of Pharmacology, University of Michigan, Ann Arbor, MI 48109, USA
contact email	avsmrcka@med.umich.edu
lab head
Naincy Chandan
contact affiliation	University of Michigan
contact email	nchandan@umich.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/2022/08/PXD027905

PRIDE project URI

• PRIDE
  1. PXD027905
     1. Label: PRIDE project
     2. Name: A Network G Protein Gai Signaling Partners is Revealed by Proximity Labeling Proteomics and Includes PDZ-RhoGEF