STEPHEN HILL's Outputs (42)

Ligand-Directed Labeling of the Adenosine A1 Receptor in Living Cells (2024)
Journal Article
Comeo, E., Goulding, J., Lin, C.-Y., Groenen, M., Woolard, J., Kindon, N. D., Harwood, C. R., Platt, S., Briddon, S. J., Kilpatrick, L. E., Scammells, P. J., Hill, S. J., & Kellam, B. (2024). Ligand-Directed Labeling of the Adenosine A1 Receptor in Living Cells. Journal of Medicinal Chemistry, 67(14), 12099–12117. https://doi.org/10.1021/acs.jmedchem.4c00835

The study of protein function and dynamics in their native cellular environment is essential for progressing fundamental science. To overcome the requirement of genetic modification of the protein or the limitations of dissociable fluorescent ligands... Read More about Ligand-Directed Labeling of the Adenosine A1 Receptor in Living Cells.

A novel and selective fluorescent ligand for the study of adenosine A2B receptors (2024)
Journal Article
Patera, F., Mistry, S. J., Kindon, N. D., Comeo, E., Gouding, J., Kellam, B., Kilpatrick, L. E., Franks, H., & Hill, S. J. (2024). A novel and selective fluorescent ligand for the study of adenosine A2B receptors. Pharmacology Research and Perspectives, 12(4), Article e1223. https://doi.org/10.1002/prp2.1223

Fluorescent ligands have proved to be powerful tools in the study of G protein-coupled receptors in living cells. Here we have characterised a new fluorescent ligand PSB603-BY630 that has high selectivity for the human adenosine A2B receptor (A2BR).... Read More about A novel and selective fluorescent ligand for the study of adenosine A2B receptors.

Design, Synthesis, and Evaluation of a New Chemotype Fluorescent Ligand for the P2Y2 Receptor (2024)
Journal Article
Knight, R., Kilpatrick, L. E., Hill, S. J., & Stocks, M. J. (2024). Design, Synthesis, and Evaluation of a New Chemotype Fluorescent Ligand for the P2Y2 Receptor. ACS Medicinal Chemistry Letters, 15(7), 1127-1135. https://doi.org/10.1021/acsmedchemlett.4c00211

The P2Y2 receptor (P2Y2R) is a target for diseases including cancer, idiopathic pulmonary fibrosis, and atherosclerosis. However, there are insufficient P2Y2R antagonists available for validating P2Y2R function and future drug development. Evaluation... Read More about Design, Synthesis, and Evaluation of a New Chemotype Fluorescent Ligand for the P2Y2 Receptor.

Mechano-sensitivity of β2-adrenoceptors enhances constitutive activation of cAMP generation that is inhibited by inverse agonists (2024)
Journal Article
Cullum, S. A., Platt, S., Dale, N., Isaac, O. C., Wragg, E. S., Soave, M., Veprintsev, D. B., Woolard, J., Kilpatrick, L. E., & Hill, S. J. (2024). Mechano-sensitivity of β2-adrenoceptors enhances constitutive activation of cAMP generation that is inhibited by inverse agonists. Communications Biology, 7(1), Article 417. https://doi.org/10.1038/s42003-024-06128-2

The concept of agonist-independent signalling that can be attenuated by inverse agonists is a fundamental element of the cubic ternary complex model of G protein-coupled receptor (GPCR) activation. This model shows how a GPCR can exist in two conform... Read More about Mechano-sensitivity of β2-adrenoceptors enhances constitutive activation of cAMP generation that is inhibited by inverse agonists.

CXCL17 is an allosteric inhibitor of CXCR4 through a mechanism of action involving glycosaminoglycans (2024)
Journal Article
White, C. W., Platt, S., Kilpatrick, L. E., Dale, N., Abhayawardana, R. S., Dekkers, S., …Hill, S. J. (2024). CXCL17 is an allosteric inhibitor of CXCR4 through a mechanism of action involving glycosaminoglycans. Science Signaling, 17(828), Article abl3758. https://doi.org/10.1126/scisignal.abl3758

CXCL17 is a chemokine principally expressed by mucosal tissues, where it facilitates chemotaxis of monocytes, dendritic cells, and macrophages and has antimicrobial properties. CXCL17 is also implicated in the pathology of inflammatory disorders and... Read More about CXCL17 is an allosteric inhibitor of CXCR4 through a mechanism of action involving glycosaminoglycans.

Kinetic analysis of fluorescent ligand binding to cell surface receptors: Insights into conformational changes and allosterism in living cells (2023)
Journal Article
Hill, S. J., & Kilpatrick, L. E. (2023). Kinetic analysis of fluorescent ligand binding to cell surface receptors: Insights into conformational changes and allosterism in living cells. British Journal of Pharmacology, https://doi.org/10.1111/bph.16185

Equilibrium binding assays are one of the mainstays of current drug discovery efforts to evaluate the interaction of drugs with receptors in membranes and intact cells. However, in recent years, there has been increased focus on the kinetics of the d... Read More about Kinetic analysis of fluorescent ligand binding to cell surface receptors: Insights into conformational changes and allosterism in living cells.

Probing expression of E-selectin using CRISPR-Cas9-mediated tagging with HiBiT in human endothelial cells (2023)
Journal Article
Ogrodzinski, L., Platt, S., Goulding, J., Alexander, C., Farr, T. D., Woolard, J., …Kilpatrick, L. E. (2023). Probing expression of E-selectin using CRISPR-Cas9-mediated tagging with HiBiT in human endothelial cells. iScience, 26(7), Article 107232. https://doi.org/10.1016/j.isci.2023.107232

E-selectin is expressed on endothelial cells in response to inflammatory cytokines and mediates leukocyte rolling and extravasation. However, studies have been hampered by lack of experimental approaches to monitor expression in real time in living c... Read More about Probing expression of E-selectin using CRISPR-Cas9-mediated tagging with HiBiT in human endothelial cells.

Characterisation of IL-23 receptor antagonists and disease relevant mutants using fluorescent probes (2023)
Journal Article
Lay, C. S., Isidro-Llobet, A., Kilpatrick, L. E., Craggs, P. D., & Hill, S. J. (2023). Characterisation of IL-23 receptor antagonists and disease relevant mutants using fluorescent probes. Nature Communications, 14, Article 2882. https://doi.org/10.1038/s41467-023-38541-2

Association of single nucleotide polymorphisms in the IL-23 receptor with several auto-inflammatory diseases, led to the heterodimeric receptor and its cytokine-ligand IL-23, becoming important drug targets. Successful antibody-based therapies direct... Read More about Characterisation of IL-23 receptor antagonists and disease relevant mutants using fluorescent probes.

Plasma membrane preassociation drives β-arrestin coupling to receptors and activation (2023)
Journal Article
Grimes, J., Koszegi, Z., Lanoiselée, Y., Miljus, T., O’Brien, S. L., Stepniewski, T. M., …Calebiro, D. (2023). Plasma membrane preassociation drives β-arrestin coupling to receptors and activation. Cell, 186(10), 2238-2255. https://doi.org/10.1016/j.cell.2023.04.018

β-arrestin plays a key role in G protein-coupled receptor (GPCR) signaling and desensitization. Despite recent structural advances, the mechanisms that govern receptor-β-arrestin interactions at the plasma membrane of living cells remain elusive. Her... Read More about Plasma membrane preassociation drives β-arrestin coupling to receptors and activation.

Use of NanoBiT and NanoBRET to characterise interleukin-23 receptor dimer formation in living cells (2022)
Journal Article
Lay, C. S., Kilpatrick, L. E., Craggs, P. D., & Hill, S. J. (2023). Use of NanoBiT and NanoBRET to characterise interleukin-23 receptor dimer formation in living cells. British Journal of Pharmacology, 180(11), 1444-1459. https://doi.org/10.1111/bph.16018

Background and Purpose: Interleukin-23 (IL-23) and its receptor are important drug targets for the treatment of auto-inflammatory diseases. IL-23 binds to a receptor complex composed of two single transmembrane spanning proteins IL23R and IL12Rβ1. In... Read More about Use of NanoBiT and NanoBRET to characterise interleukin-23 receptor dimer formation in living cells.

Kinetic analysis of endogenous β2-adrenoceptor-mediated cAMP GloSensorTM responses in HEK293 cells (2022)
Journal Article
Cullum, S. A., Veprintsev, D. B., & Hill, S. J. (2023). Kinetic analysis of endogenous β2-adrenoceptor-mediated cAMP GloSensorTM responses in HEK293 cells. British Journal of Pharmacology, 180(10), 1304-1315. https://doi.org/10.1111/bph.16008

Background and Aim: Standard pharmacological analysis of agonist activity utilises measurements of receptor-mediated responses at a set time-point, or at the peak response level, to characterise ligands. However, the occurrence of non-equilibrium con... Read More about Kinetic analysis of endogenous β2-adrenoceptor-mediated cAMP GloSensorTM responses in HEK293 cells.

Optimization of Peptide Linker-Based Fluorescent Ligands for the Histamine H1 Receptor (2022)
Journal Article
Kok, Z. Y., Stoddart, L. A., Mistry, S. J., Mocking, T. A., Vischer, H. F., Leurs, R., …Kellam, B. (2022). Optimization of Peptide Linker-Based Fluorescent Ligands for the Histamine H1 Receptor. Journal of Medicinal Chemistry, 65(12), 8258–8288. https://doi.org/10.1021/acs.jmedchem.2c00125

The histamine H1 receptor (H1R) has recently been implicated in mediating cell proliferation and cancer progression, therefore high affinity H1R-selective fluorescent ligands are desirable tools for further investigation of this behaviour in vitro an... Read More about Optimization of Peptide Linker-Based Fluorescent Ligands for the Histamine H1 Receptor.

Community guidelines for GPCR ligand bias: IUPHAR review 32 (2022)
Journal Article
Kolb, P., Kenakin, T., Alexander, S. P. H., Bermudez, M., Bohn, L. M., Breinholt, C. S., …Gloriam, D. E. (2022). Community guidelines for GPCR ligand bias: IUPHAR review 32. British Journal of Pharmacology, 179(14), 3651-3674. https://doi.org/10.1111/bph.15811

GPCRs modulate a plethora of physiological processes and mediate the effects of one-third of FDA-approved drugs. Depending on which ligand activates a receptor, it can engage different intracellular transducers. This ‘biased signalling’ paradigm requ... Read More about Community guidelines for GPCR ligand bias: IUPHAR review 32.

Role of the renin–angiotensin–aldosterone and kinin–kallikrein systems in the cardiovascular complications of COVID-19 and long COVID (2021)
Journal Article
Cooper, S. L., Boyle, E., Jefferson, S. R., Heslop, C. R. A., Mohan, P., Mohanraj, G. G. J., …Woolard, J. (2021). Role of the renin–angiotensin–aldosterone and kinin–kallikrein systems in the cardiovascular complications of COVID-19 and long COVID. International Journal of Molecular Sciences, 22(15), Article 8255. https://doi.org/10.3390/ijms22158255

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is the virus responsible for the COVID-19 pandemic. Patients may present as asymptomatic or demonstrate mild to severe and life-threatening symptoms. Although COVID-19 has a respiratory foc... Read More about Role of the renin–angiotensin–aldosterone and kinin–kallikrein systems in the cardiovascular complications of COVID-19 and long COVID.

Probing the binding of interleukin-23 to individual receptor components and the IL-23 heteromeric receptor complex in living cells using NanoBRET (2021)
Journal Article
Lay, C. S., Bridges, A., Goulding, J., Briddon, S. J., Soloviev, Z., Craggs, P. D., & Hill, S. J. (2022). Probing the binding of interleukin-23 to individual receptor components and the IL-23 heteromeric receptor complex in living cells using NanoBRET. Cell Chemical Biology, 29(1), 19-29.e6. https://doi.org/10.1016/j.chembiol.2021.05.002

Interleukin-23 (IL-23) is a pro-inflammatory cytokine involved in the host defence against pathogens, but also implicated in the development of several autoimmune disorders. The IL- 23 receptor has become a key target for drug discovery but the exact... Read More about Probing the binding of interleukin-23 to individual receptor components and the IL-23 heteromeric receptor complex in living cells using NanoBRET.

Use of NanoBiT and NanoBRET to monitor fluorescent VEGF-A binding kinetics to VEGFR2/NRP1 heteromeric complexes in living cells (2021)
Journal Article
Peach, C. J., Kilpatrick, L. E., Woolard, J., & Hill, S. J. (2021). Use of NanoBiT and NanoBRET to monitor fluorescent VEGF-A binding kinetics to VEGFR2/NRP1 heteromeric complexes in living cells. British Journal of Pharmacology, 178(12), 2393-2411. https://doi.org/10.1111/bph.15426

Background and Purpose:
VEGF‐A is a key mediator of angiogenesis, primarily signalling via VEGF receptor 2 (VEGFR2). Endothelial cells also express the co‐receptor neuropilin‐1 (NRP1) that potentiates VEGF‐A/VEGFR2 signalling. VEGFR2 and NRP1 had d... Read More about Use of NanoBiT and NanoBRET to monitor fluorescent VEGF-A binding kinetics to VEGFR2/NRP1 heteromeric complexes in living cells.

Efficient G protein coupling is not required for agonist‐mediated internalization and membrane reorganization of the adenosine A 3 receptor (2021)
Journal Article
Stoddart, L. A., Kilpatrick, L. E., Corriden, R., Kellam, B., Briddon, S. J., & Hill, S. J. (2021). Efficient G protein coupling is not required for agonist‐mediated internalization and membrane reorganization of the adenosine A 3 receptor. FASEB Journal, 35(4), Article e21211. https://doi.org/10.1096/fj.202001729rr

Organization of G protein-coupled receptors at the plasma membrane has been the focus of much recent attention. Advanced microscopy techniques have shown that these receptors can be localized to discrete microdomains and reorganization upon ligand ac... Read More about Efficient G protein coupling is not required for agonist‐mediated internalization and membrane reorganization of the adenosine A 3 receptor.

Detection of genome-edited and endogenously expressed G protein-coupled receptors (2021)
Journal Article
Soave, M., Stoddart, L. A., White, C. W., Kilpatrick, L. E., Goulding, J., Briddon, S. J., & Hill, S. J. (2021). Detection of genome-edited and endogenously expressed G protein-coupled receptors. FEBS Journal, 288(8), 2585-2601. https://doi.org/10.1111/febs.15729

© 2021 The Authors. The FEBS Journal published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies. G protein-coupled receptors (GPCRs) are the largest family of membrane receptors and major targets for FDA-approved d... Read More about Detection of genome-edited and endogenously expressed G protein-coupled receptors.

Investigation of Receptor Heteromers Using NanoBRET Ligand Binding (2021)
Journal Article
Johnstone, E. K. M., See, H. B., Abhayawardana, R. S., Song, A., Rosengren, K. J., Hill, S. J., & Pfleger, K. D. G. (2021). Investigation of Receptor Heteromers Using NanoBRET Ligand Binding. International Journal of Molecular Sciences, 22(3), Article 1082. https://doi.org/10.3390/ijms22031082

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 pharmacolog... Read More about Investigation of Receptor Heteromers Using NanoBRET Ligand Binding.

A lipid-anchored neurokinin 1 receptor antagonist prolongs pain relief by a three-pronged mechanism of action targeting the receptor at the plasma membrane and in endosomes (2021)
Journal Article
Mai, Q. N., Shenoy, P., Quach, T., Retamal, J. S., Gondin, A. B., Yeatman, H. R., …Veldhuis, N. A. (2021). A lipid-anchored neurokinin 1 receptor antagonist prolongs pain relief by a three-pronged mechanism of action targeting the receptor at the plasma membrane and in endosomes. Journal of Biological Chemistry, 296, Article 100345. https://doi.org/10.1016/J.JBC.2021.100345

G-protein-coupled receptors (GPCRs) are traditionally known for signaling at the plasma membrane, but they can also signal from endosomes after internalization to control important pathophysiological processes. In spinal neurons, sustained endosomal... Read More about A lipid-anchored neurokinin 1 receptor antagonist prolongs pain relief by a three-pronged mechanism of action targeting the receptor at the plasma membrane and in endosomes.

A nanoluciferase biosensor to investigate endogenous chemokine secretion and receptor binding (2020)
Journal Article
White, C. W., Kilpatrick, L. E., Pfleger, K. D., & Hill, S. J. (2021). A nanoluciferase biosensor to investigate endogenous chemokine secretion and receptor binding. iScience, 24(1), Article 102011. https://doi.org/10.1016/j.isci.2020.102011

© 2020 The Author(s) Secreted chemokines are critical mediators of cellular communication that elicit intracellular signaling by binding membrane-bound receptors. Here we demonstrate the development and use of a sensitive real-time approach to quanti... Read More about A nanoluciferase biosensor to investigate endogenous chemokine secretion and receptor binding.

Ligand-directed covalent labelling of a GPCR with a fluorescent tag in live cells (2020)
Journal Article
Stoddart, L. A., Kindon, N. D., Otun, O., Harwood, C. R., Patera, F., Veprintsev, D. B., …Kellam, B. (2020). Ligand-directed covalent labelling of a GPCR with a fluorescent tag in live cells. Communications Biology, 3(1), Article 722. https://doi.org/10.1038/s42003-020-01451-w

© 2020, The Author(s). To study the localisation of G protein-coupled receptors (GPCR) in their native cellular environment requires their visualisation through fluorescent labelling. To overcome the requirement for genetic modification of the recept... Read More about Ligand-directed covalent labelling of a GPCR with a fluorescent tag in live cells.

CRISPR/Cas9-mediated generation and analysis of N terminus polymorphic models of β2AR in isogenic hPSC-derived cardiomyocytes (2020)
Journal Article
Kondrashov, A., Mohd Yusof, N. A., Hasan, A., Goulding, J., Kodagoda, T., Hoang, D. M., …Denning, C. (2021). CRISPR/Cas9-mediated generation and analysis of N terminus polymorphic models of β2AR in isogenic hPSC-derived cardiomyocytes. Molecular Therapy - Methods and Clinical Development, 20, 39-53. https://doi.org/10.1016/j.omtm.2020.10.019

© 2020 During normal- and patho-physiological situations, the behavior of the beta2-adrenoreceptor (β2AR) is influenced by polymorphic variants. The functional impact of such polymorphisms has been suggested from data derived from genetic association... Read More about CRISPR/Cas9-mediated generation and analysis of N terminus polymorphic models of β2AR in isogenic hPSC-derived cardiomyocytes.

Transactivation of G protein-coupled receptors (GPCRs) and receptor tyrosine kinases (RTKs): Recent insights using luminescence and fluorescence technologies (2020)
Journal Article
Kilpatrick, L. E., & Hill, S. J. (2021). Transactivation of G protein-coupled receptors (GPCRs) and receptor tyrosine kinases (RTKs): Recent insights using luminescence and fluorescence technologies. Current Opinion in Endocrine and Metabolic Research, 16, 102-112. https://doi.org/10.1016/j.coemr.2020.10.003

© 2020 The Authors Alterations in signalling due to bidirectional transactivation of G protein-coupled receptor (GPCRs) and receptor tyrosine kinases (RTKs) are well established. Transactivation significantly diversifies signalling networks within a... Read More about Transactivation of G protein-coupled receptors (GPCRs) and receptor tyrosine kinases (RTKs): Recent insights using luminescence and fluorescence technologies.

Using Esterase Selectivity to Determine the in Vivo Duration of Systemic Availability and Abolish Systemic Side Effects of Topical β-Blockers (2020)
Journal Article
Baker, J. G., Fromont, C., Bruder, M., Thompson, K. S., Kellam, B., Hill, S. J., …Fischer, P. M. (2020). Using Esterase Selectivity to Determine the in Vivo Duration of Systemic Availability and Abolish Systemic Side Effects of Topical β-Blockers. ACS Pharmacology & Translational Science, 3(4), 737-748. https://doi.org/10.1021/acsptsci.0c00051

© 2020 American Chemical Society. For disorders of the skin, eyes, ears, and respiratory tract, topical drugs, delivered directly to the target organ, are a therapeutic option. Compared with systemic oral therapy, the benefits of topical treatments i... Read More about Using Esterase Selectivity to Determine the in Vivo Duration of Systemic Availability and Abolish Systemic Side Effects of Topical β-Blockers.

Monitoring Allosteric Interactions with CXCR4 Using NanoBiT Conjugated Nanobodies (2020)
Journal Article
Soave, M., Heukers, R., Kellam, B., Woolard, J., Smit, M. J., Briddon, S. J., & Hill, S. J. (2020). Monitoring Allosteric Interactions with CXCR4 Using NanoBiT Conjugated Nanobodies. Cell Chemical Biology, 27, 1-12. https://doi.org/10.1016/j.chembiol.2020.06.006

© 2020 The Authors Camelid single-domain antibody fragments (nanobodies) offer the specificity of an antibody in a single 15-kDa immunoglobulin domain. Their small size allows for easy genetic manipulation of the nanobody sequence to incorporate prot... Read More about Monitoring Allosteric Interactions with CXCR4 Using NanoBiT Conjugated Nanobodies.

CRISPR-Mediated Protein Tagging with Nanoluciferase to Investigate Native Chemokine Receptor Function and Conformational Changes (2020)
Journal Article
White, C. W., Caspar, B., Vanyai, H. K., Pfleger, K. D., & Hill, S. J. (2020). CRISPR-Mediated Protein Tagging with Nanoluciferase to Investigate Native Chemokine Receptor Function and Conformational Changes. Cell Chemical Biology, 27(5), 499-510. https://doi.org/10.1016/j.chembiol.2020.01.010

© 2020 The Authors G protein-coupled receptors are a major class of membrane receptors that mediate physiological and pathophysiological cellular signaling. Many aspects of receptor activation and signaling can be investigated using genetically encod... Read More about CRISPR-Mediated Protein Tagging with Nanoluciferase to Investigate Native Chemokine Receptor Function and Conformational Changes.

The effect of two selective A1-receptor agonists and the bitopic ligand VCP746 on heart rate and regional vascular conductance in conscious rats (2019)
Journal Article
Cooper, S. L., March, J., Sabbatini, A. R., Hill, S. J., Jörg, M., Scammells, P. J., & Woolard, J. (2020). The effect of two selective A1-receptor agonists and the bitopic ligand VCP746 on heart rate and regional vascular conductance in conscious rats. British Journal of Pharmacology, 177(2), 346-359. https://doi.org/10.1111/bph.14870

Background and purpose
Adenosine is a local mediator that regulates physiological and pathological processes via activation of four G protein‐coupled receptors (A1, A2A, A2B, A3). We have investigated the effect of two A1‐receptor selective agonists... Read More about The effect of two selective A1-receptor agonists and the bitopic ligand VCP746 on heart rate and regional vascular conductance in conscious rats.

Modulators of CXCR4 and CXCR7/ACKR3 Function (2019)
Journal Article
Adlere, I., Caspar, B., Arimont, M., Dekkers, S., Visser, K., Stuijt, J., …Leurs, R. (2019). Modulators of CXCR4 and CXCR7/ACKR3 Function. Molecular Pharmacology, 96(6), 737-752. https://doi.org/10.1124/mol.119.117663

Copyright © 2019 by The Author(s). The two G protein-coupled receptors (GPCRs) C-X-C chemokine receptor type 4 (CXCR4) and atypical chemokine receptor 3 (ACKR3) are part of the class A chemokine GPCR family and represent important drug targets for hu... Read More about Modulators of CXCR4 and CXCR7/ACKR3 Function.

Comparison of the ligand‐binding properties of fluorescent VEGF‐A isoforms to VEGF receptor 2 in living cells and membrane preparations using NanoBRET (2019)
Journal Article
Peach, C. J., Kilpatrick, L. E., Woolard, J., & Hill, S. J. (2019). Comparison of the ligand‐binding properties of fluorescent VEGF‐A isoforms to VEGF receptor 2 in living cells and membrane preparations using NanoBRET. British Journal of Pharmacology, 176(17), 3220-3235. https://doi.org/10.1111/bph.14755

Background and Purpose: Vascular Endothelial Growth Factor A (VEGF-A) is a key mediator of angiogenesis. A striking feature of the binding of a fluorescent analogue of VEGF165a to NanoLuciferase-tagged VEGF Receptor 2 (VEGFR2) in living cells is that... Read More about Comparison of the ligand‐binding properties of fluorescent VEGF‐A isoforms to VEGF receptor 2 in living cells and membrane preparations using NanoBRET.

Binding kinetics of ligands acting at GPCRs (2019)
Journal Article
Sykes, D. A., Stoddart, L. A., Kilpatrick, L. E., & Hill, S. J. (2019). Binding kinetics of ligands acting at GPCRs. Molecular and Cellular Endocrinology, 485, 9-19. https://doi.org/10.1016/j.mce.2019.01.018

The influence of drug-receptor binding kinetics has often been overlooked during the development of new therapeutics that target G protein-coupled receptors (GPCRs). Over the last decade there has been a growing understanding that an in-depth knowled... Read More about Binding kinetics of ligands acting at GPCRs.

Real-Time Ligand Binding of Fluorescent VEGF-A Isoforms that Discriminate between VEGFR2 and NRP1 in Living Cells (2018)
Journal Article
Peach, C. J., Kilpatrick, L. E., Friedman-Ohana, R., Zimmerman, K., Robers, M. B., Wood, K. V., …Hill, S. J. (2018). Real-Time Ligand Binding of Fluorescent VEGF-A Isoforms that Discriminate between VEGFR2 and NRP1 in Living Cells. Cell Chemical Biology, 25(10), 1208-1218.e5. https://doi.org/10.1016/j.chembiol.2018.06.012

© 2018 The Author(s) Fluorescent VEGF-A isoforms have been evaluated for their ability to discriminate between VEGFR2 and NRP1 in real-time ligand binding studies in live cells using BRET. To enable this, we synthesized single-site (N-terminal cystei... Read More about Real-Time Ligand Binding of Fluorescent VEGF-A Isoforms that Discriminate between VEGFR2 and NRP1 in Living Cells.

Characterisation of endogenous A2A and A2B receptor-mediated cyclic AMP responses in HEK 293 cells using the GloSensor™ biosensor: evidence for an allosteric mechanism of action for the A2B-selective antagonist PSB 603 (2017)
Journal Article
Goulding, J., May, L. T., & Hill, S. J. (2018). Characterisation of endogenous A2A and A2B receptor-mediated cyclic AMP responses in HEK 293 cells using the GloSensor™ biosensor: evidence for an allosteric mechanism of action for the A2B-selective antagonist PSB 603. Biochemical Pharmacology, 147, https://doi.org/10.1016/j.bcp.2017.10.013

Endogenous adenosine A2B receptors (A2BAR) mediate cAMP accumulation in HEK 293 cells. Here we have used a biosensor to investigate the mechanism of action of the A2BAR antagonist PSB 603 in HEK 293 cells. The A2A agonist CGS 21680 elicited a small r... Read More about Characterisation of endogenous A2A and A2B receptor-mediated cyclic AMP responses in HEK 293 cells using the GloSensor™ biosensor: evidence for an allosteric mechanism of action for the A2B-selective antagonist PSB 603.

Structure-activity relationships of the sustained effects of adenosine A2A receptor agonists driven by slow dissociation kinetics (2017)
Journal Article
Hothersall, J. D., Guo, D., Sarda, S., Sheppard, R. J., Chen, H., Keur, W., …Rawlins, P. B. (2017). Structure-activity relationships of the sustained effects of adenosine A2A receptor agonists driven by slow dissociation kinetics. Molecular Pharmacology, 91(1), https://doi.org/10.1124/mol.116.105551

The duration of action of adenosine A2A receptor (A2A) agonists is critical for their clinical efficacy, and we sought to better understand how this can be optimized. The in vitro temporal response profiles of a panel of A2A agonists were studied usi... Read More about Structure-activity relationships of the sustained effects of adenosine A2A receptor agonists driven by slow dissociation kinetics.

The use of fluorescence correlation spectroscopy to characterize the molecular mobility of fluorescently labelled G protein-coupled receptors (2016)
Journal Article
Kilpatrick, L. E., & Hill, S. J. (in press). The use of fluorescence correlation spectroscopy to characterize the molecular mobility of fluorescently labelled G protein-coupled receptors. Biochemical Society Transactions, 44(2), https://doi.org/10.1042/BST20150285

The membranes of living cells have been shown to be highly organized into distinct microdomains, which has spatial and temporal consequences for the interaction of membrane bound receptors and their signalling partners as complexes. Fluorescence corr... Read More about The use of fluorescence correlation spectroscopy to characterize the molecular mobility of fluorescently labelled G protein-coupled receptors.

Fragment-Based Discovery of Subtype-Selective Adenosine Receptor Ligands from Homology Models (2015)
Journal Article
Ranganathan, A., Stoddart, L. A., Hill, S. J., & Carlsson, J. (2015). Fragment-Based Discovery of Subtype-Selective Adenosine Receptor Ligands from Homology Models. Journal of Medicinal Chemistry, 58(24), 9578-9590. https://doi.org/10.1021/acs.jmedchem.5b01120

Fragment-based lead discovery (FBLD) holds great promise for drug discovery, but applications to G protein-coupled receptors (GPCRs) have been limited by a lack of sensitive screening techniques and scarce structural information. If virtual screening... Read More about Fragment-Based Discovery of Subtype-Selective Adenosine Receptor Ligands from Homology Models.

Kinetic analysis of antagonist-occupied adenosine-A3 receptors within membrane microdomains of individual cells provides evidence of receptor dimerization and allosterism (2014)
Journal Article
Corriden, R., Kilpatrick, L. E., Kellam, B., Briddon, S. J., & Hill, S. J. (2014). Kinetic analysis of antagonist-occupied adenosine-A3 receptors within membrane microdomains of individual cells provides evidence of receptor dimerization and allosterism. FASEB Journal, 28(10), 4211-4222. https://doi.org/10.1096/fj.13-247270

© The Author(s). In our previous work, using a fluorescent adenosine-A3 receptor (A3AR) agonist and fluorescence correlation spectroscopy (FCS), we demonstrated highaffinity labeling of the active receptor (R∗) conformation. In the current study, we... Read More about Kinetic analysis of antagonist-occupied adenosine-A3 receptors within membrane microdomains of individual cells provides evidence of receptor dimerization and allosterism.

The evolving small-molecule fluorescent-conjugate toolbox for Class A GPCRs (2014)
Journal Article
Vernall, A. J., Hill, S. J., & Kellam, B. (2014). The evolving small-molecule fluorescent-conjugate toolbox for Class A GPCRs. British Journal of Pharmacology, 171(5), https://doi.org/10.1111/bph.12265

The past decade has witnessed fluorescently tagged drug molecules gaining significant attraction in their use as pharmacological tools with which to visualize and interrogate receptor targets at the single-cell level. Additionally, one can generate d... Read More about The evolving small-molecule fluorescent-conjugate toolbox for Class A GPCRs.

Allosteric interactions at adenosine A1 and A3 receptors: new insights into the role of small molecules and receptor dimerization (2014)
Journal Article
Hill, S. J., May, L. T., Kellam, B., & Woolard, J. (2014). Allosteric interactions at adenosine A1 and A3 receptors: new insights into the role of small molecules and receptor dimerization. British Journal of Pharmacology, 171(5), https://doi.org/10.1111/bph.12345

Keywords:adenosine;allosterism;receptor;GPCR;dimerization;biased signalling
The purine nucleoside adenosine is present in all cells in tightly regulated concentrations. It is released under a variety of physiological and pathophysiological condition... Read More about Allosteric interactions at adenosine A1 and A3 receptors: new insights into the role of small molecules and receptor dimerization.

Monoclonal anti-β1-adrenergic receptor antibodies activate G protein signaling in the absence of β-arrestin recruitment (2013)
Journal Article
Hutchings, C. J., Cseke, G., Osborne, G., Woolard, J., Zhukov, A., Koglin, M., …Marshall, F. H. (2014). Monoclonal anti-β1-adrenergic receptor antibodies activate G protein signaling in the absence of β-arrestin recruitment. mAbs, 6(1), 246-261. https://doi.org/10.4161/mabs.27226

Thermostabilized G protein-coupled receptors used as antigens for in vivo immunization have resulted in the generation of functional agonistic anti-β1-adrenergic (β1AR) receptor monoclonal antibodies (mAbs). The focus of this study was to examine the... Read More about Monoclonal anti-β1-adrenergic receptor antibodies activate G protein signaling in the absence of β-arrestin recruitment.

Conversion of a non-selective adenosine receptor antagonist into A 3-selective high affinity fluorescent probes using peptide-based linkers (2013)
Journal Article
Vernall, A. J., Stoddart, L. A., Briddon, S. J., Ng, H. W., Laughton, C. A., Doughty, S. W., …Kellam, B. (2013). Conversion of a non-selective adenosine receptor antagonist into A 3-selective high affinity fluorescent probes using peptide-based linkers. Organic and Biomolecular Chemistry, 11(34), 5673-5682. https://doi.org/10.1039/c3ob41221k

Advances in fluorescence-based imaging technologies have helped propel the study of real-time biological readouts and analysis across many different areas. In particular the use of fluorescent ligands as chemical tools to study proteins such as G pro... Read More about Conversion of a non-selective adenosine receptor antagonist into A 3-selective high affinity fluorescent probes using peptide-based linkers.

Adenosine-A3 receptors in neutrophil microdomains promote the formation of bacteria-tethering cytonemes (2013)
Journal Article
Corriden, R., Self, T., Akong-Moore, K., Nizet, V., Kellam, B., Briddon, S. J., & Hill, S. J. (2013). Adenosine-A3 receptors in neutrophil microdomains promote the formation of bacteria-tethering cytonemes. EMBO Reports, 14(8), https://doi.org/10.1038/embor.2013.89

The A3‐adenosine receptor (A3AR) has recently emerged as a key regulator of neutrophil behaviour. Using a fluorescent A3AR ligand, we show that A3ARs aggregate in highly polarized immunomodulatory microdomains on human neutrophil membranes. In additi... Read More about Adenosine-A3 receptors in neutrophil microdomains promote the formation of bacteria-tethering cytonemes.