A5076737326- Receptor Mechanisms and Signaling
- Neuropeptides and Animal Physiology
- Neuroscience and Neuropharmacology Research
- Pharmacological Effects and Assays
- Chemical Synthesis and Analysis
- Mass Spectrometry Techniques and Applications
- Cancer, Stress, Anesthesia, and Immune Response
- Protein Kinase Regulation and GTPase Signaling
- Neuroendocrine regulation and behavior
- Phosphodiesterase function and regulation
- Nitric Oxide and Endothelin Effects
- Mitochondrial Function and Pathology
- Computational Drug Discovery Methods
- Gene Regulatory Network Analysis
- Cardiac Fibrosis and Remodeling
- Pain Mechanisms and Treatments
- Ion Transport and Channel Regulation
- Hypothalamic control of reproductive hormones
- Neurotransmitter Receptor Influence on Behavior
- Diabetes Treatment and Management
- Tryptophan and brain disorders
- Caveolin-1 and cellular processes
- Pancreatic function and diabetes
- Stress Responses and Cortisol
- Anesthesia and Neurotoxicity Research
Istituto Superiore di Sanità
2011-2024
University of Rome Tor Vergata
1992
University of Cagliari
1991
Abstract Discovering biased agonists requires a method that can reliably distinguish the bias in signalling due to unbalanced activation of diverse transduction proteins from differential amplification inherent system being studied, which invariably results non-linear nature biological networks and their measurement. We have systematically compared performance seven methods diagnostics, all are based on analysis concentration-response curves ligands according classical receptor theory....
Nociceptin/orphanin FQ (N/OFQ) controls several biological functions by selectively activating an opioid like receptor named N/OFQ peptide (NOP). Biased agonism is emerging as important and therapeutically relevant pharmacological concept in the field of G protein coupled receptors including opioids. To evaluate relevance this phenomenon NOP receptor, we used a bioluminescence resonance energy transfer technology to measure interactions with either proteins or β-arrestin 2 absence presence...
The functional selectivity of adrenergic ligands for activation β1- and β2-AR (adrenoceptor) subtypes has been extensively studied in cAMP signalling. Much less is known about ligand arrestin-mediated signalling pathways. In the present study we used resonance energy transfer methods to compare ability β2-ARs form a complex with G-protein β-subunit or β-arrestin-2 response variety agonists various degrees efficacy. profiles β1-/β2-AR two receptor-transducer interactions were sharply...
Fusion proteins between heptahelical receptors (GPCR) and G protein α-subunits show enhanced signaling efficiency in transfected cells. This is believed to be the result of molecular proximity, because interaction linked modules one chain, if not constrained by structure, should strongly favored compared with same which partners react as free species. To test this assumption we made a series fusion (type 1 4 opioid Go β2adrenergic dopamine GsL) some mutated analogs carrying different tags...
We engineered single and multiple mutations of serines 203, 204, 207 in the fifth transmembrane domain beta(2)-adrenergic receptor, a region known to interact with hydroxyl groups catechol ring. Using such mutants, we measured binding affinities panel six catecholamine agonists differing only presence substituents ethanolamine tail molecule. Although all ligands shared an intact ring, they exhibited different losses energy response mutations. For mutations, found clear relationship between...
Peptide welding technology (PWT) is a novel chemical strategy that allows the synthesis of multibranched peptides with high yield, purity and reproducibility. Using this technique, we have synthesized pharmacologically characterized tetrabranched derivatives tachykinins, substance P (SP), neurokinin A (NKA) B (NKB).The following in vitro assays were used: calcium mobilization cells expressing human recombinant NK receptors, BRET studies G-protein - NK1 receptor interaction, guinea pig ileum...
We compared the changes in binding energy generated by two mutations that shift divergent directions constitutive activity of human β 2 adrenergic receptor (β AR). A constitutively activating mutant (CAM) and double alanine replacement (AA mutant) catechol‐binding serines (S204A, S207A) helix 5 were stably expressed CHO cell lines, used to measure affinities more than 40 ligands. Moreover, efficacy same group compounds was determined as intrinsic for maximal adenylyl cyclase stimulation...
BACKGROUND AND PURPOSE Cell cycle regulators are regarded as essential for cardiomyocyte hypertrophic growth. Given that the β-adrenoceptor antagonist propranolol blunts growth, we determined whether alters expression of cell cycle-related genes in mouse hearts subjected to pressure overload. EXPERIMENTAL APPROACH Pressure overload was induced by transverse aortic constriction (TAC), whereas levels 84 were assayed real-time PCR. Propranolol (80 mg·kg−1·day−1) administered drinking water 14...
We have previously shown that myogenesis induction by Arg8-vasopressin (AVP) in L6 rat myoblasts involves a sustained stimulation of type 4 cAMP-phosphodiesterase. In this model, we observed transient cAMP generation occurs the minutes following AVP addition. Evidence suggests is due to prostaglandins produced response binding V1a receptors and subsequent activation phospholipase A2. The early increase was effective activating cAMP-dependent protein kinase (PKA) increasing phosphorylation...
We converted Ser-207, located in helix 5 of the β2-adrenergic receptor, into all other natural amino acids. To quantify receptor activation as a number-independent parameter and directly related to Gs activation, we expressed mutants Gαs-tethered form. GTP exchange such constructs is restricted fused α-subunit linear function concentration. Except S207R, were suitable level for investigation. All mutations reduced binding affinities catechol agonists, epinephrine isoproterenol, extent...
Abstract Vasopressin receptor 2 (V2R) mutations causing the nephrogenic syndrome of inappropriate antidiuresis (NSIAD) can generate two constitutively active phenotypes. One type results from residue substitutions in several V2R domains and is sensitive to vaptan inverse agonists. The other only caused by Arg 137 replacements resistant. We compared constitutive agonist-driven interactions vaptan-sensitive F229V vaptan-resistant R137C/L with β-arrestin 1, 2, Gαs, using null fibroblasts...
β<sub>2</sub>-Adrenoceptor-mediated activation of G<sub>s</sub> and adenylyl cyclase or other receptor-mediated G protein activations is believed to occur by receptor-catalyzed replacement GDP with GTP on the α-subunit protein. Here we showed that a β<sub>2</sub>-adrenoceptor-G<sub>s</sub> system, heterologously expressed in cyc<sup>-</sup> human embryonic kidney (HEK)-293 cells, can be activated presence its phosphorylation-resistant analog, guanosine 5′-<i>O</i>-(2-thiodiphosphate)...
Adrenergic receptors (AR) belong to the G protein-coupled receptor superfamily and regulate migration proliferation in various cell types. The objective of this study was evaluate whether β-AR stimulation affects antiproliferative action α2-AR agonists on B16F10 cells and, if so, determine relative contribution subtypes. Using pharmacological approaches, evaluation Ki-67 expression by flow cytometry luciferase-based cAMP assay, we found that treatment with isoproterenol, a agonist, increased...
Altered β-adrenergic receptor (β-AR) density has been reported in cells, animals, and humans receiving β-blocker treatment. In some cases, β-AR is upregulated, but others, it unaffected or even reduced. Collectively, these results would imply that changes β-blockade are not related. However, still clarified whether the effects of β-blockers on related to their ability activate different signaling pathways. To this aim, five clinically relevant endowed with inverse, partial biased agonism at...