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Subtype selective fluorescent ligands based on ICI 118,551 to study the human β2‐adrenoceptor in CRISPR/Cas9 genome‐edited HEK293T cells at low expression levels (2021)
Journal Article
Kellam, B., White, C. W., Goulding, J., Mistry, S. J., Soave, M., Woolard, J., …Hill, S. J. (2021). Subtype selective fluorescent ligands based on ICI 118,551 to study the human β2‐adrenoceptor in CRISPR/Cas9 genome‐edited HEK293T cells at low expression levels. Pharmacology Research and Perspectives, 9(3), Article e00779. https://doi.org/10.1002/prp2.779

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 β2-adrenocepto... Read More about Subtype selective fluorescent ligands based on ICI 118,551 to study the human β2‐adrenoceptor in CRISPR/Cas9 genome‐edited HEK293T cells at low expression levels.

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.

Development and Application of Subtype-Selective Fluorescent Antagonists for the Study of the Human Adenosine A1 Receptor in Living Cells (2021)
Journal Article
Comeo, E., Trinh, P., Nguyen, A. T., Nowell, C. J., Kindon, N. D., Soave, M., …Scammells, P. J. (2021). Development and Application of Subtype-Selective Fluorescent Antagonists for the Study of the Human Adenosine A1 Receptor in Living Cells. Journal of Medicinal Chemistry, 64(10), 6670-6695. https://doi.org/10.1021/acs.jmedchem.0c02067

The adenosine A1 receptor (A1AR) is a G-protein-coupled receptor (GPCR) that provides important therapeutic opportunities for a number of conditions including congestive heart failure, tachycardia, and neuropathic pain. The development of A1AR-select... Read More about Development and Application of Subtype-Selective Fluorescent Antagonists for the Study of the Human Adenosine A1 Receptor in Living Cells.

The use of fluorescence correlation spectroscopy to monitor cell surface ?2?adrenoceptors at low expression levels in human embryonic stem cell?derived cardiomyocytes and fibroblasts (2021)
Journal Article
Goulding, J., Kondrashov, A., Mistry, S. J., Melarangi, T., Vo, N. T. N., Hoang, D. M., …Hill, S. J. (2021). The use of fluorescence correlation spectroscopy to monitor cell surface β2‐adrenoceptors at low expression levels in human embryonic stem cell‐derived cardiomyocytes and fibroblasts. FASEB Journal, 35(4), Article e21398. https://doi.org/10.1096/fj.202002268r

The importance of cell phenotype in determining the molecular mechanisms underlying ?2- adrenoceptor (?2AR) function has been noted previously when comparing responses in primary cells and recombinant model cell lines. Here, we have generated haploty... Read More about The use of fluorescence correlation spectroscopy to monitor cell surface ?2?adrenoceptors at low expression levels in human embryonic stem cell?derived cardiomyocytes and fibroblasts.

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 ?2AR 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 ?2AR 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 ?-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 ?-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.

Subtype-Selective Fluorescent Ligands as Pharmacological Research Tools for the Human Adenosine A2A Receptor (2019)
Journal Article
Comeo, E., Kindon, N. D., Soave, M., Stoddart, L. A., Kilpatrick, L. E., Scammells, P. J., …Kellam, B. (2020). Subtype-Selective Fluorescent Ligands as Pharmacological Research Tools for the Human Adenosine A2A Receptor. Journal of Medicinal Chemistry, 63(5), 2656-2672. https://doi.org/10.1021/acs.jmedchem.9b01856

© 2019 American Chemical Society. Among class A G protein-coupled receptors (GPCR), the human adenosine A2A receptor (hA2AAR) remains an attractive drug target. However, translation of A2AAR ligands into the clinic has proved challenging and an impro... Read More about Subtype-Selective Fluorescent Ligands as Pharmacological Research Tools for the Human Adenosine A2A Receptor.

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.

NanoBiT Complementation to Monitor Agonist-Induced Adenosine A1 Receptor Internalization (2019)
Journal Article
Soave, M., Kellam, B., Woolard, J., Briddon, S. J., & Hill, S. J. (2019). NanoBiT Complementation to Monitor Agonist-Induced Adenosine A1 Receptor Internalization. Slas Discovery, https://doi.org/10.1177/2472555219880475

Receptor internalization in response to prolonged agonist treatment is an important regulator of G protein–coupled receptor (GPCR) function. The adenosine A1 receptor (A1AR) is one of the adenosine receptor family of GPCRs, and evidence for its agoni... Read More about NanoBiT Complementation to Monitor Agonist-Induced Adenosine A1 Receptor Internalization.

Optimised insert design for improved single-molecule imaging and quantification through CRISPR-Cas9 mediated knock-in (2019)
Journal Article
Khan, A. O., White, C. W., Pike, J. A., Yule, J., Slater, A., Hill, S. J., …Morgan, N. V. (2019). Optimised insert design for improved single-molecule imaging and quantification through CRISPR-Cas9 mediated knock-in. Scientific Reports, 9(1), Article 14219. https://doi.org/10.1038/s41598-019-50733-9

© 2019, The Author(s). 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 sys... Read More about Optimised insert design for improved single-molecule imaging and quantification through CRISPR-Cas9 mediated knock-in.

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.