ORCID Profile
0000-0002-4424-239X
Current Organisation
University of Nottingham
Does something not look right? The information on this page has been harvested from data sources that may not be up to date. We continue to work with information providers to improve coverage and quality. To report an issue, use the Feedback Form.
Publisher: Wiley
Date: 07-1998
Publisher: Wiley
Date: 05-11-2015
DOI: 10.1111/BPH.13316
Publisher: Wiley
Date: 02-2010
Publisher: American Society for Pharmacology & Experimental Therapeutics (ASPET)
Date: 19-01-2010
Publisher: Wiley
Date: 29-06-2011
DOI: 10.1096/FJ.11-186296
Publisher: Wiley
Date: 31-03-2010
Publisher: American Chemical Society (ACS)
Date: 16-03-2021
Publisher: Elsevier BV
Date: 2008
DOI: 10.1016/J.BIOCEL.2008.04.001
Abstract: G protein-coupled receptors (GPCRs) are a major target in the drug discovery process. One important response that results from activation of a wide range of GPCRs is activation of the ERK signalling cascade. Given the abundance of both upstream activators and downstream targets of ERK1/2, the precise spatiotemporal control of ERK1/2 phosphorylation is crucial for maintaining the specificity of the physiological outcome. ERK activity is regulated via a number of mechanisms including compartmentalisation and scaffolding proteins. These scaffolding proteins can enhance the transduction of a specific signalling pathway by targeting pathway components to particular intracellular locations or signalling complexes. Recently, a number of fluorescent indicators of ERK1/2 phosphorylation have been developed that allow the regulation of this pathway to be investigated with greater spatiotemporal resolution than was previously possible. These fluorescent probes in conjunction with those for other signalling cascades should help unravel the spatiotemporal organisation of this pathway.
Publisher: American Chemical Society (ACS)
Date: 14-06-2021
Publisher: Wiley
Date: 05-1999
Publisher: Elsevier BV
Date: 2018
Publisher: Wiley
Date: 10-1998
Publisher: Springer Science and Business Media LLC
Date: 06-2015
DOI: 10.1038/NMETH.3398
Publisher: Wiley
Date: 06-02-2020
DOI: 10.1111/BPH.14575
Publisher: Cold Spring Harbor Laboratory
Date: 30-11-2018
DOI: 10.1101/482596
Abstract: The use of CRISPR-Cas9 genome editing to introduce endogenously expressed tags has the potential to address a number of the classical limitations of single molecule localisation microscopy. In this work we present the first systematic comparison of inserts introduced through CRISPR- knock in, with the aim of optimising this approach for single molecule imaging. We show that more highly monomeric and codon optimised variants of mEos result in improved expression at the TubA1B locus, despite the use of identical guides, homology templates, and selection strategies. We apply this approach to target the G protein-coupled receptor (GPCR) CXCR4 and show a further insert dependent effect on expression and protein function. Finally, we show that compared to over-expressed CXCR4, endogenously labelled s les allow for accurate single molecule quantification on ligand treatment. This suggests that despite the complications evident in CRISPR mediated labelling, the development of CRISPR-PALM has substantial quantitative benefits.
Publisher: Wiley
Date: 05-2021
DOI: 10.1002/PRP2.779
Abstract: Fluorescent ligand technologies have proved to be powerful tools to improve our understanding of ligand‐receptor interactions. Here we have characterized a small focused library of nine fluorescent ligands based on the highly selective β 2 ‐adrenoceptor (β 2 AR) antagonist ICI 118,551. The majority of fluorescent ICI 118,551 analogs had good affinity for the β 2 AR (pK D .0) with good selectivity over the β 1 AR (pK D .0). The most potent and selective ligands being 8c (ICI 118,551‐Gly‐Ala‐BODIPY‐FL‐X β 2 AR pK D 7.48), 9c (ICI 118,551‐βAla‐βAla‐BODIPY‐FL‐X β 2 AR pK D 7.48), 12a (ICI 118,551‐PEG‐BODIPY‐X‐630/650 β 2 AR pK D 7.56), and 12b (ICI 118,551‐PEG‐BODIPY‐FL β 2 AR pK D 7.42). 9a (ICI 118,551‐βAla‐βAla‐BODIPY‐X‐630/650) had the highest affinity at recombinant β 2 ARs (pK D 7.57), but also exhibited significant binding affinity to the β 1 AR (pK D 6.69). Nevertheless, among the red fluorescent ligands, 9a had the best imaging characteristics in recombinant HEK293 T cells and labeling was mostly confined to the cell surface. In contrast, 12a showed the highest propensity to label intracellular β 2 ARs in HEK293 T cell expressing exogenous β 2 ARs. This suggests that a combination of the polyethylene glycol (PEG) linker and the BODIPY‐X‐630/650 makes this ICI 118,551 derivative particularly susceptible to crossing the cell membrane to access the intracellular β 2 ARs. We have also used these ligands in combination with CRISPR/Cas9 genome‐edited HEK293 T cells to undertake for the first time real‐time ligand binding to native HEK293 T β 2 ARs at low native receptor expression levels. These studies provided quantitative data on ligand‐binding characteristics but also allowed real‐time visualization of the ligand‐binding interactions in genome‐edited cells using NanoBRET luminescence imaging.
Publisher: Royal Society of Chemistry (RSC)
Date: 2018
DOI: 10.1039/C7RA13148H
Abstract: Mild, metal free aromatization of tetrahydroisoquinolinols. Synthesis of (benzimidazolyl)isoquinolinols.
Publisher: The Company of Biologists
Date: 15-02-2012
DOI: 10.1242/JCS.091090
Abstract: The central and pervasive influence of cAMP on cellular functions underscores the value of stringent control of the organization of adenylyl cyclases (ACs) in the plasma membrane. Biochemical data suggest that ACs reside in membrane rafts and could compartmentalize intermediary scaffolding proteins and associated regulatory elements. However, little is known about the organization or regulation of the dynamic behaviour of ACs in a cellular context. The present study examines these issues, using confocal image analysis of various AC8 constructs, combined with fluorescence recovery after photobleaching and fluorescence correlation spectroscopy. These studies reveal that AC8, through its N-terminus, enhances the cortical actin signal at the plasma membrane an interaction that was confirmed by GST pull-down and immunoprecipitation experiments. AC8 also associates dynamically with lipid rafts the direct association of AC8 with sterols was confirmed in Förster resonance energy transfer experiments. Disruption of the actin cytoskeleton and lipid rafts indicates that AC8 tracks along the cytoskeleton in a cholesterol-enriched domain, and the cAMP that it produces contributes to sculpting the actin cytoskeleton. Thus, an adenylyl cyclase is shown not just to act as a scaffold, but also to actively orchestrate its own micro-environment, by associating with the cytoskeleton and controlling the association by producing cAMP, to yield a highly organized signalling hub.
Publisher: American Society for Pharmacology & Experimental Therapeutics (ASPET)
Date: 22-06-2010
Publisher: Elsevier BV
Date: 05-2023
Publisher: Wiley
Date: 12-2041
Publisher: Elsevier BV
Date: 04-2011
Publisher: Elsevier BV
Date: 05-2020
Publisher: Elsevier BV
Date: 2021
Publisher: Wiley
Date: 11-1997
Publisher: Wiley
Date: 27-02-0027
DOI: 10.1111/BPH.12345
Publisher: MDPI AG
Date: 22-01-2021
DOI: 10.3390/IJMS22031082
Abstract: Receptor heteromerization is the formation of a complex involving at least two different receptors with pharmacology that is distinct from that exhibited by its constituent receptor units. Detection of these complexes and monitoring their pharmacology is crucial for understanding how receptors function. The Receptor-Heteromer Investigation Technology (Receptor-HIT) utilizes ligand-dependent modulation of interactions between receptors and specific biomolecules for the detection and profiling of heteromer complexes. Previously, the interacting biomolecules used in Receptor-HIT assays have been intracellular proteins, however in this study we have for the first time used bioluminescence resonance energy transfer (BRET) with fluorescently-labeled ligands to investigate heteromerization of receptors on the cell surface. Using the Receptor-HIT ligand binding assay with NanoBRET, we have successfully investigated heteromers between the angiotensin II type 1 (AT1) receptor and the β2 adrenergic receptor (AT1-β2AR heteromer), as well as between the AT1 and angiotensin II type 2 receptor (AT1-AT2 heteromer).
Location: United Kingdom of Great Britain and Northern Ireland
Location: United Kingdom of Great Britain and Northern Ireland
Location: United Kingdom of Great Britain and Northern Ireland
No related grants have been discovered for Stephen Hill.