A5007776580- Receptor Mechanisms and Signaling
- Neuropeptides and Animal Physiology
- Free Radicals and Antioxidants
- Sleep and Wakefulness Research
- Neuroscience and Neuropharmacology Research
- Chemical Synthesis and Analysis
- Monoclonal and Polyclonal Antibodies Research
- Mass Spectrometry Techniques and Applications
- Molecular spectroscopy and chirality
- Diabetes Treatment and Management
- Eicosanoids and Hypertension Pharmacology
- Computational Drug Discovery Methods
- Pharmacological Effects and Assays
- Adenosine and Purinergic Signaling
- Ion Transport and Channel Regulation
- Chemokine receptors and signaling
- Enzyme Structure and Function
- Protein Kinase Regulation and GTPase Signaling
- Cell death mechanisms and regulation
- T-cell and B-cell Immunology
- Crystallography and molecular interactions
- Adrenal Hormones and Disorders
- Inflammasome and immune disorders
- Biochemical effects in animals
- Crystallization and Solubility Studies
Washington University in St. Louis
2023
Heptares Therapeutics (United Kingdom)
2013-2022
University of Hertfordshire
2014
University of Michigan–Ann Arbor
2003
Fragment screening of a thermostabilized mGlu5 receptor using high-concentration radioligand binding assay enabled the identification moderate affinity, high ligand efficiency (LE) pyrimidine hit 5. Subsequent optimization structure-based drug discovery methods led to selection 25, HTL14242, as an advanced lead compound for further development. Structures stabilized complexed with 25 and another molecule in series, 14, were determined at resolutions 2.6 3.1 Å, respectively.
The adenosine A(2A) receptor (A(2A)R) is a G-protein-coupled that plays key role in transmembrane signalling mediated by the agonist adenosine. structure of A(2A)R was determined recently an antagonist-bound conformation, which facilitated T4 lysozyme fusion cytoplasmic loop 3 and considerable stabilisation conferred on bound inverse ZM241385. Unfortunately, natural does not sufficiently stabilise for formation diffraction-quality crystals. As first step towards determining to agonist,...
Abstract G-protein-coupled receptors signal through cognate G proteins. Despite the widespread importance of these receptors, their regulatory mechanisms for G-protein selectivity are not fully understood. Here we present a native mass spectrometry-based approach to interrogate both biased signalling and allosteric modulation β 1 -adrenergic receptor in response various ligands. By simultaneously capturing effects ligand binding coupling different proteins, probed relative specific...
The orexin system, which consists of the two G protein-coupled receptors OX1 and OX2, activated by neuropeptides OX-A OX-B, is firmly established as a key regulator behavioral arousal, sleep, wakefulness has been an area intense research effort over past decades. X-ray structures in complex with 10 new antagonist ligands from diverse chemotypes are presented, complement existing structural information for system highlight critical importance lipophilic hotspots water molecules these...
Abstract Potassium‐coupled chloride transporters (KCCs) play crucial roles in regulating cell volume and intracellular concentration. They are characteristically inhibited under isotonic conditions via phospho‐regulatory sites located within the cytoplasmic termini. Decreased inhibitory phosphorylation response to hypotonic swelling stimulates transport activity, dysfunction of this regulatory process has been associated with various human diseases. Here, we present cryo‐EM structures KCC3b...
Native mass spectrometry (MS) is proving to be a disruptive technique for studying the interactions of proteins, necessary understanding functional roles these biomolecules. Recent research expanding application native MS towards membrane proteins directly from isolated preparations or purified detergent micelles. The former results in complex spectra comprising several heterogeneous protein complexes; latter enables therapeutic targets screened against multiplexed compound libraries. In...
Thermostabilized G protein-coupled receptors used as antigens for in vivo immunization have resulted the generation of functional agonistic anti-β1-adrenergic (β1AR) receptor monoclonal antibodies (mAbs). The focus this study was to examine pharmacology these evaluate their mechanistic activity at β1AR. Immunization with β1AR stabilized yielded five stable hybridoma clones, four which expressed IgG, determined cell-based assays cAMP stimulation. bind diverse epitopes associated low nanomolar...
Abstract G Protein Coupled Receptors (GPCRs) constitute the largest family of signalling proteins responsible for translating extracellular stimuli into intracellular functions. They play crucial roles in numerous physiological processes and are major targets drug discovery. Dysregulation GPCRs is implicated various diseases, making understanding their structural dynamics critical therapeutic development. Here, we use Hydrogen Deuterium Exchange Mass Spectrometry (HDX-MS) to explore turkey...
ADGRL4/ELTD1 is an orphan adhesion GPCR (aGPCR) expressed in endothelial cells that regulates tumour angiogenesis. The majority of aGPCRs are receptors. Stachel Hypothesis proposes a mechanism for aGPCR activation, which contain tethered agonist (termed Stachel) C-terminal to the GPCR-proteolytic site (GPS) cleavage point which, when exposed, initiates canonical signalling. This has been shown growing number aGPCRs. We tested this hypothesis on by designing full length (FL) and fragment...
Recent technical advances have greatly facilitated G-protein coupled receptors crystallography as evidenced by the number of successful x-ray structures that been reported recently. These include novel detergents, specialised techniques well protein engineering solutions such fusions and conformational thermostabilisation. Using thermostabilisation, it is possible to generate variants GPCRs exhibit significantly increased stability in detergent micelles whilst preferentially occupying a...
Abstract G-protein coupled receptors (GPCRs) are important therapeutic targets since more than 40% of the drugs on market exert their action through these proteins. To decipher molecular mechanisms activation and signaling, GPCRs often need to be isolated reconstituted from a detergent-solubilized state into well-defined controllable lipid model system. Several methods exist reconstitute membrane proteins in systems but usually reconstitution success is tested at end experiment by an...
Gastrin-deficient mice have impaired basal and agonist-stimulated gastric acid secretion. To analyze whether an intrinsic parietal cell defect contributed to the reduced secretion, we analyzed calcium responses secretory function in vitro. Parietal cells were purified by light-scatter sorting gastrin, histamine, carbachol measured gastrin-deficient wild-type preparations. Surprisingly, histamine-induced concentrations higher mutant [(14)C]aminopyrine uptake analysis acutely isolated glands...