A5066088240- Receptor Mechanisms and Signaling
- Neuropeptides and Animal Physiology
- Mass Spectrometry Techniques and Applications
- Pancreatic function and diabetes
- Protein Structure and Dynamics
- Monoclonal and Polyclonal Antibodies Research
- Bacterial Genetics and Biotechnology
- Advanced Proteomics Techniques and Applications
- Neuroendocrine regulation and behavior
- RNA and protein synthesis mechanisms
- Protein Kinase Regulation and GTPase Signaling
- Microbial Metabolic Engineering and Bioproduction
- Chemical Synthesis and Analysis
- Chemokine receptors and signaling
- Computational Drug Discovery Methods
- Lipid Membrane Structure and Behavior
- Neuroscience and Neuropharmacology Research
- Wastewater Treatment and Nitrogen Removal
- Peptidase Inhibition and Analysis
- Cannabis and Cannabinoid Research
- Diabetes Treatment and Management
- Mitochondrial Function and Pathology
- Electrolyte and hormonal disorders
- Cellular transport and secretion
- Amino Acid Enzymes and Metabolism
Philipps University of Marburg
2024
Stanford University
2020-2023
Institute for Research in Immunology and Cancer
2016-2023
Université de Montréal
2016-2023
MRC Laboratory of Molecular Biology
2021-2023
Paul Scherrer Institute
2013-2022
Max Perutz Labs
2022
University of Vienna
2022
Tohoku University
2022
Queen's University Belfast
2022
Abstract Two-thirds of human hormones and one-third clinical drugs activate ~350 G-protein-coupled receptors (GPCR) belonging to four classes: A, B1, C F. Whereas a model activation has been described for class very little is known about the other classes, which differ by being activated endogenous ligands bound mainly or entirely extracellularly. Here we show that, although they use same structural scaffold share several ‘helix macroswitches’, GPCR classes in their ‘residue microswitch’...
Abstract G-protein-coupled receptors do not only feature the orthosteric pockets, where most endogenous agonists bind, but also a multitude of other allosteric pockets that have come into focus as potential binding sites for synthetic modulators. Here, to better characterise such we investigate 557 GPCR structures by exhaustively docking small molecular probes in silico and converting ensemble locations pocket-defining volumes. Our analysis confirms all previously identified reveals nine...
Abstract In addition to G protein-coupled receptor (GPCR) desensitization and endocytosis, β-arrestin recruitment ligand-stimulated GPCRs promotes non-canonical signalling cascades. Distinguishing the respective contributions of β-arrestin-promoted endocytosis in propagating has been limited by lack selective analytical tools. Here, using a combination virtual screening cell-based assays, we have identified small molecule that selectively inhibits interaction between β2-adaptin subunit...
Binding of arrestin to phosphorylated G protein-coupled receptors (GPCRs) is crucial for modulating signaling. Once internalized, some GPCRs remain complexed with β-arrestins, while others interact only transiently; this difference affects GPCR signaling and recycling. Cell-based in vitro biophysical assays reveal the role membrane phosphoinositides (PIPs) β-arrestin recruitment GPCR-β-arrestin complex dynamics. We find that broadly stratify into two groups, one requires PIP binding does...
ABSTRACT Nitrogen regulation in Escherichia coli is a model system for gene bacteria. Growth on glutamine as sole nitrogen source assumed to be limiting, inferred from slow growth and strong NtrB/NtrC-dependent activation. However, we show that under these conditions, the intracellular concentration not limiting but 5.6-fold higher than ammonium-replete conditions; addition, α-ketoglutarate concentrations are elevated. We address this paradox systems perspective. dominant role of NtrC...
Site-directed scanning mutagenesis is a powerful protein engineering technique which allows studies of functionality at single amino acid resolution and design stabilized proteins for structural biophysical work. However, creating libraries hundreds mutants remains challenging, expensive time-consuming process. The efficiency the step key fast economical generation such libraries. PCR artefacts as misannealing tandem primer repeats are often observed in cloning reduce mutagenesis. Here we...
Heterotrimeric guanine nucleotide-binding protein (G protein)-coupled receptors (GPCRs) bind to extracellular ligands and drugs modulate intracellular responses through conformational changes. Despite their importance as drug targets, the molecular origins of pharmacological properties such efficacy (maximum signaling response) potency (the ligand concentration at half-maximal remain poorly understood for any ligand-receptor-signaling system. We used prototypical adrenaline-β2 adrenergic...
Scanning mutagenesis is a powerful protein engineering technique used to study structure-function relationship, map binding sites and design more stable proteins or with altered properties. One of the time-consuming tasks encountered in application this primers for site-directed mutagenesis. Here we present an open-source multi-platform software AAscan developed task according set empirical rules such as melting temperature, overall length, length overlap regions, presence GC clamps at 3'...
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...
Ammonium assimilation in Escherichia coli is regulated by two paralogous proteins (GlnB and GlnK), which orchestrate interactions with regulators of gene expression, transport proteins, metabolic pathways. Yet how they conjointly modulate the activity glutamine synthetase, key enzyme for nitrogen assimilation, poorly understood. We combine experiments theory to study dynamic roles GlnB GlnK during starvation upshift. measure time-resolved vivo concentrations metabolites, total...
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...
Activation of G protein-coupled receptors by agonists may result in the activation one or more proteins and recruitment arrestins. The extent each these pathways depends on intrinsic efficacy ligand. Quantification relative to a reference compound is essential for development novel compounds. In operational model, changes can be compensated "functional" affinity, resulting poorly defined values. To separate effects ligand affinity from activity receptor, we developed Michaelis-Menten based...
Abstract Cannabinoid CB1 and CB2 receptors are members of the G protein-coupled receptor family, which is largest class membrane proteins in human genome. As part endocannabinoid system, they have many regulatory functions body. Their malfunction therefore triggers a diverse set undesired conditions, such as pain, neuropathy, nephropathy, pruritus, osteoporosis, cachexia Alzheimer’s disease. Although drugs targeting system exist, molecular functional mechanisms involved still poorly...
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...
Summary Binding of arrestin to phosphorylated G protein-coupled receptors (GPCRs) is crucial for modulating signaling. Once internalized some GPCRs may complex with arrestin, while others interact transiently; this difference affects receptor signaling and recycling. Cell-based in vitro biophysical assays reveal the role membrane phosphoinositides (PIPs) recruitment GPCR-arrestin dynamics. We find that broadly stratify into two groups, one requiring PIP-binding does not. Plasma PIPs...
Site-directed scanning mutagenesis is a useful tool applied in studying protein function and designing proteins with new properties, such as increased stability or enzymatic activity. Creating systematic library of hundreds site-directed mutants still demanding expensive task. The established protocols for making libraries include PCR amplification the recombinant DNA using pair primers carrying target mutation same PCR. Unfortunately, this approach very often coupled artifacts which...
G protein-coupled receptors are important therapeutic drug targets for a wide range of diseases. Their ability to preferentially engage specific signaling pathways over others can be exploited design drugs that target only disease-associated leading an improved safety profile. However, the underlying molecular mechanisms preferential pathway engagement complex and remain largely elusive. To elucidate multifaceted actions at receptor level lead coupling, we employ combination techniques. Our...
Abstract Ammonium assimilation in E. coli is regulated by two paralogous proteins (GlnB and GlnK), which orchestrate interactions with regulators of gene expression, transport metabolic pathways. Yet how they conjointly modulate the activity glutamine synthetase (GS), key enzyme for nitrogen assimilation, poorly understood. We combine experiments theory to study dynamic roles GlnB GlnK during starvation upshift. measure time-resolved vivo concentrations metabolites, total...
Abstract G protein-coupled receptors (GPCRs) drive an array of critical physiological functions and are important class drug targets, though a map which GPCR genetic variants associated with phenotypic variation is lacking. We performed phenome-wide association analysis for 269 common protein-altering in 156 GPCRs 275 phenotypes, including disease outcomes diverse quantitative measurements, using 337,205 UK Biobank participants identified 138 associations. discovered novel associations...