A5039325764- Receptor Mechanisms and Signaling
- Protein Kinase Regulation and GTPase Signaling
- Neuropeptides and Animal Physiology
- Computational Drug Discovery Methods
- Pancreatic function and diabetes
- Monoclonal and Polyclonal Antibodies Research
- Lipoproteins and Cardiovascular Health
- Neuroscience and Neuropharmacology Research
- Diabetes, Cardiovascular Risks, and Lipoproteins
- Cancer, Lipids, and Metabolism
- Chemokine receptors and signaling
- Endoplasmic Reticulum Stress and Disease
- Ion channel regulation and function
- Lipid metabolism and biosynthesis
- Cholesterol and Lipid Metabolism
- Neurofibromatosis and Schwannoma Cases
- Protein Degradation and Inhibitors
- Advanced Proteomics Techniques and Applications
- Pharmacogenetics and Drug Metabolism
- Hormonal Regulation and Hypertension
- Lipid Membrane Structure and Behavior
- Viral Infectious Diseases and Gene Expression in Insects
- Adenosine and Purinergic Signaling
- Cancer, Stress, Anesthesia, and Immune Response
- Cellular transport and secretion
Institute for Research in Immunology and Cancer
2007-2024
Université de Montréal
2007-2024
Tohoku University
2017
Japan Agency for Medical Research and Development
2017
Japan Science and Technology Agency
2017
Centre hospitalier universitaire de Québec
2007
New England Center for Children
1994
Royal Victoria Hospital
1990-1991
McGill University
1990-1991
Columbia University
1990
Abstract Endocytosis and intracellular trafficking of receptors are pivotal to maintain physiological functions drug action; however, robust quantitative approaches lacking study such processes in live cells. Here we present new bioluminescence resonance energy transfer (BRET) sensors quantitatively monitor G protein-coupled (GPCRs) β-arrestin trafficking. These based on bystander BRET use the naturally interacting chromophores luciferase (RLuc) green fluorescent protein (rGFP) from Renilla...
The recognition that individual GPCRs can activate multiple signaling pathways has raised the possibility of developing drugs selectively targeting therapeutically relevant ones. This requires tools to determine which G proteins and βarrestins are activated by a given receptor. Here, we present set BRET sensors monitoring activation 12 protein subtypes based on translocation their effectors plasma membrane (EMTA). Unlike most existing detection systems, EMTA does not require modification...
Synthesis and maturation of G protein-coupled receptors are complex events that require an intricate combination processes include protein folding, post-translational modifications, transport through distinct cellular compartments. Relatively little is known about the nature kinetics specific steps involved in these processes. Here, human δ opioid receptor expressed embryonic kidney 293S cells used as a model to delineate establish synthesis, glycosylation, transport. We found synthesized...
Cholesteryl ester transfer protein (CETP) mediates an important pathway for reverse cholesterol transport. Concentrations of CETP in fasting plasma were measured by radioimmunoassay two different groups hyperlipoproteinemic subjects. Plasma concentrations correlated closely with activity normal (r = 0.86). In the first group 58 patients, significantly increased, as compared those 79 subjects and hypercholesterolemic (+26%) combined (+25%) but not altered moderately hypertriglyceridemic...
A MAb (TP-2) directed against human cholesteryl ester transfer protein (CETP) has been applied to the development of a competitive solid-phase RIA. Experiments with immobilized CETP have shown that upon incubation plasma or HDL in presence Tween (0.05%) apo A-I (but not A-II) binds while TP-2 binding is concomitantly decreased. With high detergent concentration (0.5% Triton), interference eliminated and specific RIA which all fractions same affinity can be obtained. Plasma levels CETP, A-I,...
We have previously shown that only a fraction of the newly synthesized human δ opioid receptors is able to leave endoplasmic reticulum (ER) and reach cell surface (Petäjä-Repo, U. E, Hogue, M., Laperrière, A., Walker, P., Bouvier, M. (2000) <i>J. Biol. Chem.</i> 275, 13727–13736). In present study, we investigated fate those are retained intracellularly. Pulse-chase experiments revealed disappearance receptor precursor form (<i>M</i> <sub>r</sub> 45,000) two smaller species 42,000 39,000)...
The reactivity of a series monoclonal antibodies directed against human low density lipoproteins (LDL) has been tested with hepatic and intestinal apolipoprotein B (apo-B) termed B-100 B-48, respectively (Kane, J. P., Hardman, D. A., Paulus, H. E. (1980) Proc. Natl. Acad. Sci. U. S. A. 77, 2465-2469). Whereas those that have previously shown to recognize determinants close the LDL receptor recognition site reacted only B-100, two specific for other regions apo-B both B-48. Therefore, it is...
G protein-coupled receptors are key signaling molecules and major targets for pharmaceuticals. The concept of ligand-dependent biased raises the possibility developing drugs with improved efficacy safety profiles, yet translating this to native tissues remains a challenge. Whether drug activity profiling in recombinant cell-based assays, traditionally used discovery, has any relevance physiology is unknown. Here we focused on mu opioid receptor, unrivalled target pain treatment also driver...
G protein–coupled receptors (GPCRs) regulate cellular signaling processes by coupling to diverse combinations of heterotrimeric proteins composed Gα, Gβ, and Gγ subunits. Biosensors based on bioluminescence resonance energy transfer (BRET) have advanced our understanding GPCR functional selectivity. Some BRET biosensors monitor ligand-induced conformational changes in the receptor or proteins, whereas others recruitment downstream effectors sites protein activation. Here, we compared ability...
Data in the literature suggest that circulating levels of lipoprotein(a) [Lp(a)] and insulinlike growth factor I (IGF-I) respond similarly to therapy with hormone, estrogen, or tamoxifen. To more clearly document these relations, we designed a randomized, double-blind, placebo-controlled study effects tamoxifen continuous estrogen on Lp(a), IGF-I, IGF binding protein 3 (IGFBP-3) healthy postmenopausal women. Both decreased serum IGF-I 30% below baseline during months treatment, while IGFBP-3...
The β2 adrenergic receptor (β2AR) increases intracellular Ca2+ in a variety of cell types. By combining pharmacological and genetic manipulations, we reveal novel mechanism through which the β2AR promotes mobilization (pEC50 = 7.32 ± 0.10) nonexcitable human embryonic kidney (HEK)293S cells. Downregulation Gs with sustained cholera toxin pretreatment use Gs-null HEK293 (∆Gs-HEK293) cells generated using clustered regularly interspaced short palindromic repeat-associated protein-9 nuclease...
Probucol is a hypolipidemic agent that causes marked decrease in high density lipoprotein (HDL) cholesterol. To investigate the mechanism of this effect, two studies were performed hypercholesterolemic patients who had been stabilized previously on diet and not receiving other lipid-lowering medication. Plasma cholesteryl ester transfer protein (CETP) concentrations measured fasting plasma samples before after 10 weeks probucol therapy using sensitive specific radioimmunoassay. total low...
In addition to their interactions with hetero-trimeric G proteins, seven-transmembrane domain receptors are now known form multimeric complexes that can include receptor homo- or hetero-oligomers and/or accessory proteins modulate activity. The calcitonin gene-related peptide (CGRP) requires the assembly of receptor-like single-transmembrane activity-modifying protein-1 reach cell surface and be active. However, relative stoichiometric arrangement these two within a complex remains unknown....
Protein palmitoylation is a reversible lipid modification that plays important roles for many proteins involved in signal transduction, but relatively little known about the regulation of this and cellular location where it occurs. We demonstrate human delta opioid receptor palmitoylated at two distinct locations embryonic kidney 293 cells undergoes dynamic one these sites. Although could be readily observed mature (Mr 55,000), [3H]palmitate incorporation into precursor 45,000) detected only...
This study assessed how conformational information encoded by ligand binding to <i>δ</i>-opioid receptors (DORs) is transmitted Kir3.1/Kir3.2 channels. Human embryonic kidney 293 cells were transfected with bioluminescence resonance energy transfer (BRET) donor/acceptor pairs that allowed us evaluate independently reciprocal interactions among signaling partners. These and coimmunoprecipitation studies indicated DORs, G<i>βγ</i>, Kir3 subunits constitutively interacted one another....
The expression of low density lipoprotein (LDL) antigenic determinants in the delipidated and solubilized apolipoprotein B (apo-B) free sodium dodecyl sulfate (SDS) has been studied.Of six distinct which react with previously characterized monoclonal antibodies against LDL (Milne, R. W., Theolis, R., Jr., Verdery, B., Marcel, Y. L. (1983) Arteriosclerosis 3, 23-30), only one, that recognized by antibody 1D1, was expressed on soluble apo-B, indicating apo-B may be partly denatured.The average...
G protein-coupled receptors (GPCRs) have been shown to activate the mitogen-activated protein kinases, ERK1/2, through both protein-dependent and -independent mechanisms. Here, we describe a protein-independent mechanism that unravels an unanticipated role for β-arrestins. Stimulation of V2 vasopressin receptor (V2R) in cultured cells or vivo rat kidney medullar collecting ducts led activation ERK1/2 metalloproteinase-mediated shedding factor activating insulin-like growth (IGFR). This...
A comprehensive understanding of signalling downstream GPCRs requires a broad approach to capture novel modalities in addition established pathways. Here, using an array sixteen validated BRET-based biosensors, we analyzed the ability seven different β-adrenergic ligands engage five distinct pathways β1-adrenergic receptor (β1AR). In generating signatures and capturing functional selectivity for toward these pathways, also revealed coupling that have not previously been ascribed βAR. These...
The ability of individual G protein-coupled receptors (GPCR) to engage multiple signaling pathways opens opportunities for the development better drugs. This requires new knowledge and tools determine protein subtypes barrestins engaged by a given receptor. Here, we used BRET-based effector membrane translocation assay (EMTA) that monitors activation each Gα through recruitment selective effectors βarrestins plasma membrane. Profiling 100 therapeutically relevant GPCR revealed great...
Abstract Communication across membranes controls critical cellular processes and is achieved by receptors translating extracellular signals into selective cytoplasmic responses. While receptor tertiary structures can be readily characterized, associations quaternary are challenging to study their implications in signal transduction remain poorly understood. Here, we report a computational approach for predicting self-associations, designing oligomers with various signaling properties. Using...
Biochemical and functional evidence suggest that the calcitonin receptor-like receptor (CRLR) interacts with activity-modifying protein-1 (RAMP1) to generate a gene-related peptide (CGRP) receptor. Using bioluminescence resonance energy transfer (BRET), we investigated oligomeric assembly of CRLR−RAMP1 signaling complex in living cells. As for their wild-type counterparts, fusion proteins linking CRLR RAMP1 donor Renilla luciferase (Rluc) acceptor green fluorescent protein (GFP) reach cell...
Abstract The recognition that individual GPCRs can activate multiple signaling pathways has raised the possibility of developing drugs selectively targeting therapeutically relevant ones. This requires tools to determine which G proteins and βarrestins are activated by a given receptor. Here, we present set BRET sensors monitoring activation 12 protein subtypes based on translocation their effectors plasma membrane (EMTA). Unlike most existing detection systems, EMTA does not require...