A5005097721- Receptor Mechanisms and Signaling
- Pharmacological Effects and Assays
- Lipid Membrane Structure and Behavior
- Monoclonal and Polyclonal Antibodies Research
- Computational Drug Discovery Methods
- Neuropeptides and Animal Physiology
- Adenosine and Purinergic Signaling
- Chemical Synthesis and Analysis
- Protein Structure and Dynamics
- Neuroscience and Neuropharmacology Research
- Analytical Chemistry and Chromatography
- Pharmacological Receptor Mechanisms and Effects
- Cannabis and Cannabinoid Research
- Advanced biosensing and bioanalysis techniques
- Enzyme Catalysis and Immobilization
- Carbohydrate Chemistry and Synthesis
- Mass Spectrometry Techniques and Applications
- Pancreatic function and diabetes
University of Birmingham
2020-2025
University of Nottingham
2020-2025
Queen's Medical Centre
2020-2024
University of Kent
1975
Abstract To study the localisation of G protein-coupled receptors (GPCR) in their native cellular environment requires visualisation through fluorescent labelling. overcome requirement for genetic modification receptor or limitations dissociable ligands, here we describe rational design a compound that covalently and selectively labels GPCR living cells with moiety. We designed antagonist, which linker incorporated between pharmacophore (ZM241385) fluorophore (sulfo-cyanine5) is able to...
The study of protein function and dynamics in their native cellular environment is essential for progressing fundamental science. To overcome the requirement genetic modification or limitations dissociable fluorescent ligands, ligand-directed (LD) chemistry has most recently emerged as a complementary, bioorthogonal approach labeling proteins. Here, we describe rational design, development, application first A1AR living cells. We pharmacologically demonstrate covalent expressed cells while...
Introduction The β 2 -adrenoceptor (β AR) is a class A G protein-coupled receptor (GPCR). It therapeutically relevant in asthma and chronic obstructive pulmonary disease (COPD), where AR agonists relieve bronchoconstriction. prototypical GPCR for structural biophysical studies. However, the molecular basis of agonist efficacy at not understood. We hypothesised that kinetics GPCR–G protein interactions could play role determining ligand efficacy. By studying range with varying efficacy, we...
Abstract G protein‐coupled receptors (GPCRs) are valuable therapeutic targets for many diseases. A central question of GPCR drug discovery is to understand what determines the agonism or antagonism ligands that bind them. Ligands exert their action via interactions in ligand binding pocket. We hypothesized there a common set receptor made by diverse structures mediate and among large dataset different ligands, functionally important will be over‐represented. computationally docked ~2700...
Abstract Introduction The β 2 -adrenoceptor (β AR) is a class A G protein-coupled receptor (GPCR). It therapeutically relevant in asthma, whereby AR agonists relieve bronchoconstriction. prototypical GPCR for structural and biophysical studies. However, the molecular basis of agonist efficacy at not understood. We hypothesized that kinetics ligand binding GPCR-G protein interactions could play role efficacy. characterised pharmacology range examined correlation between mini-G s Methods used...
The β2-adrenoceptor (β2AR) is a well-established target in asthma and prototypical G protein-coupled receptor for biophysical studies. Solubilization of membrane proteins has classically involved the use detergents. However, detergent environment differs from native often destabilizes proteins. Use amphiphilic copolymers promising strategy to solubilize within their lipid complete absence Here we show isolation β2AR polymer diisobutylene maleic acid (DIBMA). We demonstrate that remains...
Introduction: The cannabinoid receptor (CBR) subtypes 1 (CB1R) and 2 (CB2R) are key components of the endocannabinoid system (ECS), playing a central role in control peripheral pain, inflammation immune response, with further roles endocrine regulation food intake energy balance. So far, few medicines targeting these receptors have reached clinic, suggesting that better understanding signalling properties existing tool compounds clinical candidates may open door to development more effective...
Abstract The β2-adrenoceptor (β2AR) is a well-established target in asthma and prototypical GPCR for biophysical studies. Solubilisation of membrane proteins has classically involved the use detergents. However, detergent environment differs from native often destabilises proteins. Use amphiphilic copolymers promising strategy to solubilise within their lipid complete absence Here we show isolation β 2 AR polymer Diisobutylene maleic acid (DIBMA). We demonstrate that remains functional DIBMA...
Measurements of membrane protein thermostability reflect ligand binding. Current assays often require purification or rely on pre-existing radiolabelled fluorescent ligands, limiting their application to established targets. Alternative methods, such as fluorescence-detection size exclusion chromatography thermal shift, detect aggregation but are not amenable high-throughput screening. Here, we present a ThermoBRET method quantify the relative G coupled receptors (GPCRs), using cannabinoid (CB
Abstract Measurements of membrane protein thermostability allows indirect detection ligand binding. Current assays require purification or rely on pre-existing radiolabelled fluorescent ligands, limiting their application to established target proteins. Alternative methods detect aggregation which requires sufficiently high level expression. Here, we present a ThermoBRET method quantify the relative G coupled receptors (GPCRs), using cannabinoid (CB 1 and CB 2 ) β -adrenoceptor (β AR) as...
The β2-adrenoceptor (β2AR) is a well-established target in asthma and prototypical GPCR for biophysical studies. Solubilisation of membrane proteins has classically involved the use detergents. However, detergent environment differs from native often destabilises proteins. Use amphiphilic copolymers promising strategy to solubilise within their lipid complete absence Here we show isolation β2AR polymer Diisobutylene maleic acid (DIBMA). We demonstrate that remains functional DIBMA particle...
Abstract Here, we describe rational design of a compound that covalently and selectively labels G protein-coupled receptor (GPCR) in living cells with fluorescent moiety. Using wild-type adenosine A 2A as model system, show labelling without impeding access to the orthosteric binding site demonstrate its use endogenously expressing systems. This offers non-invasive selective approach study function localisation native GPCRs.
Abstract G protein coupled receptors (GPCRs) are valuable therapeutic targets for many diseases. A central question of GPCR drug discovery is to understand what determines the agonism or antagonism ligands which bind them. Ligands exert their action via interactions in ligand binding pocket. We hypothesised that there a common set receptor made by diverse structures mediate and among large dataset different ligands, functionally important will be over-represented. computationally docked...