Allosteric modulation of adenosine A1 and cannabinoid 1 receptor signaling by G-peptides

© 2020 The Authors. Pharmacology Research & Perspectives published by John Wiley & Sons Ltd, British Pharmacological Society and American Society for Pharmacology and Experimental Therapeutics..

While allosteric modulation of GPCR signaling has gained prominence to address the need for receptor specificity, efforts have mainly focused on allosteric sites adjacent to the orthosteric ligand-binding pocket and lipophilic molecules that target transmembrane helices. In this study, we examined the allosteric influence of native peptides derived from the C-terminus of the Gα subunit (G-peptides) on signaling from two Gi-coupled receptors, adenosine A1 receptor (A1 R) and cannabinoid receptor 1 (CB1 ). We expressed A1 R and CB1 fusions with G-peptides derived from Gαs, Gαi, and Gαq in HEK 293 cells using systematic protein affinity strength modulation (SPASM) and monitored the impact on downstream signaling in the cell compared to a construct lacking G-peptides. We used agonists N6 -Cyclopentyladenosine (CPA) and 5'-N-Ethylcarboxamidoadenosine (NECA) for A1 R and 2-Arachidonoylglycerol (2-AG) and WIN 55,212-2 mesylate (WN) for CB1 . G-peptides derived from Gαi and Gαq enhance agonist-dependent cAMP inhibition, demonstrating their effect as positive allosteric modulators of Gi-coupled signaling. In contrast, both G-peptides suppress agonist-dependent IP1 levels suggesting that they differentially function as negative allosteric modulators of Gq-coupled signaling. Taken together with our previous studies on Gs-coupled receptors, this study provides an extended model for the allosteric effects of G-peptides on GPCR signaling, and highlights their potential as probe molecules to enhance receptor specificity.

Medienart:

E-Artikel

Erscheinungsjahr:

2020

Erschienen:

2020

Enthalten in:

Zur Gesamtaufnahme - volume:8

Enthalten in:

Pharmacology research & perspectives - 8(2020), 6 vom: 06. Dez., Seite e00673

Sprache:

Englisch

Beteiligte Personen:

Touma, Anja M [VerfasserIn]
Malik, Rabia U [VerfasserIn]
Gupte, Tejas [VerfasserIn]
Sivaramakrishnan, Sivaraj [VerfasserIn]

Links:

Volltext

Themen:

Adenosine A1
Adenosine-5’-(N-ethylcarboxamide)
CB1
Cannabinoid
Chromogranins
EC 3.6.1.-
EC 3.6.5.1
GINIP protein, rat
GTP-Binding Protein alpha Subunits, Gq-G11
GTP-Binding Protein alpha Subunits, Gs
GTP-binding proteins
Gnas protein, mouse
Intracellular Signaling Peptides and Proteins
Journal Article
N(6)-cyclopentyladenosine
Peptide Fragments
Receptor
Receptor, Adenosine A1
Receptor, Cannabinoid, CB1
Research Support, N.I.H., Extramural
SCH 442 416
Signal transduction

Anmerkungen:

Date Completed 03.09.2021

Date Revised 03.09.2021

published: Print

Citation Status MEDLINE

doi:

10.1002/prp2.673

funding:

Förderinstitution / Projekttitel:

PPN (Katalog-ID):

NLM316932779