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Profiling the diversity of agonist-selective effects on the proximal proteome environment of G protein-coupled receptors

ABSTRACT: G-protein coupled receptors (GPCRs) regulate various aspects of cellular behavior and represent actionable targets for drug discovery. Activation of specific signaling pathways downstream of G-protein coupled receptors (GPCRs) or targeting the receptors at selected cellular locations has the potential to provide therapeutic actions with fewer side effects. However, the understanding of the molecular mechanisms underlying GPCR activation is limited, hampering drug discovery efforts towards selective ligands. Proximity biotin labeling based on an engineered ascorbic acid peroxidase (APEX) combined with quantitative mass spectrometry is a powerful method to delineate these mechanisms given its capacity to simultaneously capture proximal protein interaction networks and the cellular location of the receptor. However, a major challenge is to extract the various information from these complex datasets. Here, we describe a computational framework for proximity labeling datasets which predicts ligand-dependent subcellular location of GPCRs and quantitatively deconvolutes the effect of receptor location and proximal interactors. We applied this approach to the mu-opioid receptor and not only monitored distinct effects of ligands on receptor trafficking, but also discovered two novel regulators, EYA4 and KCTD12, which modulate MOR-driven G protein-dependent signaling.

ORGANISM(S): Homo Sapiens

SUBMITTER: Ruth Huttenhain

PROVIDER: PXD031393 | panorama | Sun Mar 31 00:00:00 GMT 2024

REPOSITORIES: PanoramaPublic

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Profiling the proximal proteome of the activated μ-opioid receptor.

Polacco Benjamin J BJ Lobingier Braden T BT Blythe Emily E EE Abreu Nohely N Khare Prachi P Howard Matthew K MK Gonzalez-Hernandez Alberto J AJ Xu Jiewei J Li Qiongyu Q Novy Brandon B Naing Zun Zar Chi ZZC Shoichet Brian K BK Coyote-Maestas Willow W Levitz Joshua J Krogan Nevan J NJ Von Zastrow Mark M Hüttenhain Ruth R

Nature chemical biology 20240325 9

The μ-opioid receptor (μOR) represents an important target of therapeutic and abused drugs. So far, most understanding of μOR activity has focused on a subset of known signal transducers and regulatory molecules. Yet μOR signaling is coordinated by additional proteins in the interaction network of the activated receptor, which have largely remained invisible given the lack of technologies to interrogate these networks systematically. Here we describe a proteomics and computational approach to ma ...[more]

PMID: 38528119

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Project description:Chang2008 - ERK activation, hallucinogenic drugs mediated signalling through serotonin receptors This model is described in the article: Towards a quantitative representation of the cell signaling mechanisms of hallucinogens: measurement and mathematical modeling of 5-HT1A and 5-HT2A receptor-mediated ERK1/2 activation. Chang CW, Poteet E, Schetz JA, Gümü? ZH, Weinstein H. Neuropharmacology 2009; 56 Suppl 1: 213-225 Abstract: Through a multidisciplinary approach involving experimental and computational studies, we address quantitative aspects of signaling mechanisms triggered in the cell by the receptor targets of hallucinogenic drugs, the serotonin 5-HT2A receptors. To reveal the properties of the signaling pathways, and the way in which responses elicited through these receptors alone and in combination with other serotonin receptors' subtypes (the 5-HT1AR), we developed a detailed mathematical model of receptor-mediated ERK1/2 activation in cells expressing the 5-HT1A and 5-HT2A subtypes individually, and together. In parallel, we measured experimentally the activation of ERK1/2 by the action of selective agonists on these receptors expressed in HEK293 cells. We show here that the 5-HT1AR agonist Xaliproden HCl elicited transient activation of ERK1/2 by phosphorylation, whereas 5-HT2AR activation by TCB-2 led to higher, and more sustained responses. The 5-HT2AR response dominated the MAPK signaling pathway when co-expressed with 5-HT1AR, and diminution of the response by the 5-HT2AR antagonist Ketanserin could not be rescued by the 5-HT1AR agonist. Computational simulations produced qualitative results in good agreement with these experimental data, and parameter optimization made this agreement quantitative. In silico simulation experiments suggest that the deletion of the positive regulators PKC in the 5-HT2AR pathway, or PLA2 in the combined 5-HT1A/2AR model greatly decreased the basal level of active ERK1/2. Deletion of negative regulators of MKP and PP2A in 5-HT1AR and 5-HT2AR models was found to have even stronger effects. Under various parameter sets, simulation results implied that the extent of constitutive activity in a particular tissue and the specific drug efficacy properties may determine the distinct dynamics of the 5-HT receptor-mediated ERK1/2 activation pathways. Thus, the mathematical models are useful exploratory tools in the ongoing efforts to establish a mechanistic understanding and an experimentally testable representation of hallucinogen-specific signaling in the cellular machinery, and can be refined with quantitative, function-related information. This model is hosted on BioModels Database and identified by: MODEL0975191032. To cite BioModels Database, please use: BioModels Database: An enhanced, curated and annotated resource for published quantitative kinetic models. To the extent possible under law, all copyright and related or neighbouring rights to this encoded model have been dedicated to the public domain worldwide. Please refer to CC0 Public Domain Dedication for more information.

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Dataset Information

Profiling the diversity of agonist-selective effects on the proximal proteome environment of G protein-coupled receptors

Publications

Profiling the proximal proteome of the activated μ-opioid receptor.

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