A5060489555- Receptor Mechanisms and Signaling
- GaN-based semiconductor devices and materials
- Neuropeptides and Animal Physiology
- Neurobiology and Insect Physiology Research
- Chemokine receptors and signaling
- Monoclonal and Polyclonal Antibodies Research
- Analytical Chemistry and Sensors
- Electrolyte and hormonal disorders
- Neuroendocrine regulation and behavior
- Computational Drug Discovery Methods
- bioluminescence and chemiluminescence research
- Neuroscience and Neuropharmacology Research
- Protein Kinase Regulation and GTPase Signaling
- T-cell and B-cell Immunology
- Renin-Angiotensin System Studies
- Metal and Thin Film Mechanics
- Analog and Mixed-Signal Circuit Design
- Biomedical and Engineering Education
- Quantum-Dot Cellular Automata
- Biochemical Analysis and Sensing Techniques
- Growth Hormone and Insulin-like Growth Factors
- Neuroscience and Neural Engineering
- Sleep and Wakefulness Research
- Advanced Glycation End Products research
- Pharmacological Effects and Assays
Harry Perkins Institute of Medical Research
2004-2022
The University of Western Australia
2006-2022
Australian Research Council
2021-2022
Personalis (United States)
2022
Queen Elizabeth II Medical Centre
2016-2021
Keogh Institute for Medical Research
2001
Sir Charles Gairdner Hospital
2001
The ability of G-protein-coupled receptors (GPCRs) to interact form new functional structures, either forming oligomers with themselves or associations other intracellular proteins, has important implications for the regulation cellular events; however, little is known about how this occurs. Here, we have employed a newly emerging technology, bioluminescence resonance energy transfer (BRET), used study protein-protein interactions in living cells, demonstrate that thyrotropin-releasing...
Activation of the type 1 angiotensin II receptor (AT1) triggers proinflammatory signaling through pathways independent classical Gq that regulate vascular homeostasis. Here, we report AT1 preformed a heteromeric complex with for advanced glycation endproducts (RAGE). by (Ang II) triggered transactivation cytosolic tail RAGE and NF-κB-driven gene expression independently liberation ligands or ligand-binding ectodomain RAGE. The importance this pathway was demonstrated our finding adverse...
The αvβ3 integrin is known to cooperate with receptor tyrosine kinases enhance cellular responses. To determine whether regulates transforming growth factor β (TGFβ) 1-induced responses, we investigated the interaction between and TGFβ type II (TGFβIIR) in primary human lung fibroblasts. We report that TGFβ1 up-regulates cell surface mRNA expression of a time- dose-dependent manner. Co-immunoprecipitation confocal microscopy showed TGFβRII associates clusters αvβ3, following exposure. This...
The bioluminescence resonance energy transfer (BRET) technique has become extremely popular for studying proteinprotein interactions in living cells and real time.Of particular interest is the ability to monitor between G protein-coupled receptors, such as thyrotropin-releasing hormone receptor (TRHR), proteins critical regulating their function, β-arrestin.Using TRHR/β-arrestin interactions, we have demonstrated improvements all 3 generations of BRET (BRET 1 , 2 eBRET) by using novel forms...
The Duffy antigen/receptor for chemokines (DARC) is an unusual chemokine receptor that binds a large number of inflammatory both the CC and CXC families with nanomolar affinity, yet it lacks ability to signal upon ligand binding. Using bioluminescent resonant energy transfer, we have demonstrated first time DARC exists as constitutive homo-oligomer in living cells furthermore hetero-oligomerizes CCR5. DARC-CCR5 interaction impairs chemotaxis calcium flux through CCR5, whereas internalization...
Background and Purpose The C ‐ X chemokine receptors 3 ( CXCR3 ) 4 CXCR4 are involved in various autoimmune diseases cancers. Small antagonists have previously been shown to cross‐inhibit binding , CC 2 CCR2 5 CCR5 heteromers. We investigated whether can form heteromeric complexes the characteristics of chemokines small ligand compounds these receptor Experimental Approach – heteromers were identified HEK293T cells using co‐immunoprecipitation, time‐resolved fluorescence resonance energy...
Leptin, the 16-kDa peptide hormone product of ob gene, regulates body weight via hypothalamus but also influences several aspects reproductive function. Results previous studies have suggested that pregnancy is a state leptin resistance, because food consumption remains stable or increases despite progressive rise in plasma across most gestation. In present study, we assessed whether this apparent resistance during rat was due to either increased leptin-binding activity and/or reduced...
We have provided the first evidence for specific heteromerization between α1A-adrenoceptor (α1AAR) and CXC chemokine receptor 2 (CXCR2) in live cells. α1AAR CXCR2 are both expressed areas such as stromal smooth muscle layer of prostate. By utilizing G protein-coupled (GPCR) heteromer identification technology on cell-based bioluminescence resonance energy transfer (BRET) assay platform, our studies human embryonic kidney 293 cells identified norepinephrine-dependent β-arrestin recruitment...
Nephrogenic syndrome of inappropriate antidiuresis (NSIAD) is a genetic disease first described in 2 unrelated male infants with severe symptomatic hyponatremia. Despite undetectable arginine vasopressin levels, patients have inappropriately concentrated urine resulting hyponatremia, hypoosmolality, and natriuresis. Here, we describe functionally characterize novel type receptor (V2R) gain-of-function mutation. An L312S substitution the seventh transmembrane domain was identified boy...
G-protein-coupled receptors (GPCRs) are regulated by a complex network of mechanisms such as oligomerization and internalization. Using the GPCR subtypes for thyrotropin-releasing hormone (TRHR1 TRHR2), aim this study was to determine if subtype-specific differences exist in trafficking process. If so, we wished impact homo- hetero-oligomerization on TRHR subtype potential mechanism differential cellular responses induced TRH. Expression either β-arrestin 1 or 2 promoted TRHR1 In contrast,...
Understanding the role of G protein-coupled receptor (GPCR; also known as a 7 transmembrane receptor) heteromerization in physiology and pathophysiology cellular function has now become major research focus. However, there is currently lack cell-based assays capable profiling specific functional consequences ligand-dependent manner. pharmacology specifically associated with heteromer contrast to monomer or homomer enables so-called biochemical fingerprints be ascertained. This first step...
Solubilization of mineralized bone by osteoclasts is largely dependent on the acidification extracellular resorption lacuna driven vacuolar (H+)-ATPases (V-ATPases) polarized within ruffled border membranes. V-ATPases consist two functionally and structurally distinct domains, V1 V0. The peripheral cytoplasmically oriented domain drives ATP hydrolysis, which necessitates translocation protons across integral membrane bound V0 domain. Here, we demonstrate that an accessory subunit, Ac45,...
The bioluminescence resonance energy transfer (BRET) technique has become extremely valuable for the real-time monitoring of protein-protein interactions in live cells. This method is highly amenable to detection G protein-coupled receptor (GPCR) with proteins critical regulating their function, such as β-arrestins. Of particular interest endocrinologists ability monitor involving endocrine receptors, orexin 2 or vasopressin type II receptor. BRET utilizes heterologous co-expression fusion...
Arginine vasopressin (AVP) is released from the posterior pituitary and controls water homeostasis. AVP binding to V2 receptors (V2Rs) located on kidney collecting duct epithelial cells triggers activation of Gs proteins, leading increased cAMP levels, trafficking aquaporin-2 channels, consequent permeability antidiuresis. Typically, loss-of-function V2R mutations cause nephrogenic diabetes insipidus (NDI), whereas gain-of-function syndrome inappropriate antidiuresis (NSIAD). Here we provide...
Background and Purpose The orexin system regulates a multitude of key physiological processes, particularly involving maintenance metabolic homeostasis. Consequently, there is considerable potential for pharmaceutical development the treatment disorders from narcolepsy to syndrome. It acts through hormonal activity two endogenous peptides, A binding receptors 1 2 ( OX ) with similar affinity, B higher affinity than receptors. We have previously revealed data differentiating receptor subtypes...
Heteromerization can play an important role in regulating the activation and/or signal transduction of most forms receptors, including receptor tyrosine kinases (RTKs). The study heteromerization has evolved extensively with emergence resonance energy transfer based approaches such as bioluminescence (BRET). Here, we report adaptation our Receptor-Heteromer Investigation Technology (Receptor-HIT) that recently been published G protein-coupled (GPCR) Heteromer Identification (GPCR-HIT). We...
We have investigated application of AlGaN/GaN cell-based biosensors for detection biological reactions in response to the drug ionomycin. The device sensitivity was dependent on environmental factors such as temperature and atmospheric composition. A significant improvement signal amplitude obtained through stabilisation measurement conditions. These results are an important step development optimisation reliable biosensors.
The angiotensin type 2 (AT ) receptor and the bradykinin (B are G protein-coupled receptors (GPCRs) that have major roles in cardiovascular system. two known to functionally interact at various levels, there is some evidence observed crosstalk may occur as a result of heteromerization. We investigated for heteromerization AT B HEK293FT cells using bioluminescence resonance energy transfer (BRET)-proximity based assays, including Receptor Heteromer Investigation Technology (Receptor-HIT)...
β-arrestins bind to phosphorylated, seven-transmembrane-spanning, G protein-coupled receptors (GPCRs), including the type 1 angiotensin II receptor (AT<sub>1</sub>R), promote desensitization and internalization. The AT<sub>1</sub> R is a class B GPCR that recruits both β-arrestin1 β-arrestin2, forming stable complexes cotraffic deep-core endocytic vesicles. β-Arrestins contain one amphipathic potentially amphitropic (membrane-targeting) α-helix (helix I) may translocation membrane or...
This investigation of the biocompatibility AlGaN/GaN HEMT structure and HEK-293FT cells shows that material is suitable for future biological applications. In our experiment wells were filled with HEK-293-FT cell in Dulbecco's Modified Eagle Medium (DMEM), pieces wafers placed some while others left as control comparison growth mortality rates. These samples prepared by cleaning sequentially acetone, isopropanol, deionised water. A total 48 small diameter 8mm used, where 24 wells. The sizes...