Identification of the interactome of the DP1 receptor for Prostaglandin D
DP1
GPCR
IQGAP1
Interactome
Signaling
Trafficking
Journal
Biochimica et biophysica acta. General subjects
ISSN: 1872-8006
Titre abrégé: Biochim Biophys Acta Gen Subj
Pays: Netherlands
ID NLM: 101731726
Informations de publication
Date de publication:
11 2021
11 2021
Historique:
received:
26
11
2020
revised:
19
07
2021
accepted:
25
07
2021
pubmed:
6
8
2021
medline:
17
11
2021
entrez:
5
8
2021
Statut:
ppublish
Résumé
Mechanisms governing localization, trafficking and signaling of G protein-coupled receptors (GPCRs) are critical in cell function. Protein-protein interactions are determinant in these processes. However, there are very little interacting proteins known to date for the DP1 receptor for prostaglandin D We performed LC-MS/MS analyses of the DP1 receptor interactome in HEK293 cells. To functionally validate our LC-MS/MS data, we studied the implications of the interaction with the IQGAP1 scaffold protein in the trafficking and signaling of DP1. In addition to expected interacting proteins such as heterotrimeric G protein subunits, we identified proteins involved in signaling, trafficking, and folding localized in various cell compartments. Endogenous DP1-IQGAP1 co-immunoprecipitation was observed in colon cancer HT-29 cells. The interaction was augmented by DP1 agonist activation in HEK293 cells and GST-pulldown assays showed that IQGAP1 binds to intracellular loops 2 and 3 of DP1. Co-localization of the two proteins was observed by confocal microscopy at the cell periphery and in intracellular vesicles in the basal state. PGD Our findings define the putative DP1 interactome, a patho-physiologically important receptor, and validated the interaction with IQGAP1 in DP1 function. Our data also reveal that IQGAP proteins may differentially regulate GPCR signaling. The identified putative DP1-interacting proteins open multiple lines of research in DP1 and GPCR biology in various cell compartments.
Sections du résumé
BACKGROUND
Mechanisms governing localization, trafficking and signaling of G protein-coupled receptors (GPCRs) are critical in cell function. Protein-protein interactions are determinant in these processes. However, there are very little interacting proteins known to date for the DP1 receptor for prostaglandin D
METHODS
We performed LC-MS/MS analyses of the DP1 receptor interactome in HEK293 cells. To functionally validate our LC-MS/MS data, we studied the implications of the interaction with the IQGAP1 scaffold protein in the trafficking and signaling of DP1.
RESULTS
In addition to expected interacting proteins such as heterotrimeric G protein subunits, we identified proteins involved in signaling, trafficking, and folding localized in various cell compartments. Endogenous DP1-IQGAP1 co-immunoprecipitation was observed in colon cancer HT-29 cells. The interaction was augmented by DP1 agonist activation in HEK293 cells and GST-pulldown assays showed that IQGAP1 binds to intracellular loops 2 and 3 of DP1. Co-localization of the two proteins was observed by confocal microscopy at the cell periphery and in intracellular vesicles in the basal state. PGD
CONCLUSIONS
Our findings define the putative DP1 interactome, a patho-physiologically important receptor, and validated the interaction with IQGAP1 in DP1 function. Our data also reveal that IQGAP proteins may differentially regulate GPCR signaling.
GENERAL SIGNIFICANCE
The identified putative DP1-interacting proteins open multiple lines of research in DP1 and GPCR biology in various cell compartments.
Identifiants
pubmed: 34352343
pii: S0304-4165(21)00128-8
doi: 10.1016/j.bbagen.2021.129969
pii:
doi:
Substances chimiques
IQ motif containing GTPase activating protein 1
0
Receptors, Immunologic
0
Receptors, Prostaglandin
0
ras GTPase-Activating Proteins
0
Prostaglandin D2
RXY07S6CZ2
prostaglandin D2 receptor
XZF106QU24
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
129969Subventions
Organisme : CIHR
Pays : Canada
Informations de copyright
Copyright © 2021 Elsevier B.V. All rights reserved.