Visualizing Ligand Binding to a GPCR In Vivo Using NanoBRET
Diana C. Alcobia,
Alexandra I. Ziegler,
Alexander Kondrashov,
Eleonora Comeo,
Sarah Mistry,
Barrie Kellam,
Aeson Chang,
Jeanette Woolard,
Stephen J. Hill,
Erica K. Sloan
Affiliations
Diana C. Alcobia
Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, Nottingham NG7 2UH, UK; Centre of Membrane Proteins and Receptors, University of Birmingham and University of Nottingham, The Midlands, UK; Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC 3052, Australia
Alexandra I. Ziegler
Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC 3052, Australia
Alexander Kondrashov
Wolfson Centre for Stem Cells, Tissue Engineering & Modelling (STEM), Centre for Biomolecular Sciences, University of Nottingham, Nottingham NG7 2RD, UK
Eleonora Comeo
Centre of Membrane Proteins and Receptors, University of Birmingham and University of Nottingham, The Midlands, UK; School of Pharmacy, Division of Biomolecular Science and Medicinal Chemistry, Centre for Biomolecular Sciences, University of Nottingham, Nottingham NG7 2RD, UK
Sarah Mistry
Centre of Membrane Proteins and Receptors, University of Birmingham and University of Nottingham, The Midlands, UK; School of Pharmacy, Division of Biomolecular Science and Medicinal Chemistry, Centre for Biomolecular Sciences, University of Nottingham, Nottingham NG7 2RD, UK
Barrie Kellam
Centre of Membrane Proteins and Receptors, University of Birmingham and University of Nottingham, The Midlands, UK; School of Pharmacy, Division of Biomolecular Science and Medicinal Chemistry, Centre for Biomolecular Sciences, University of Nottingham, Nottingham NG7 2RD, UK
Aeson Chang
Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC 3052, Australia
Jeanette Woolard
Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, Nottingham NG7 2UH, UK; Centre of Membrane Proteins and Receptors, University of Birmingham and University of Nottingham, The Midlands, UK; Corresponding author
Stephen J. Hill
Division of Physiology, Pharmacology and Neuroscience, School of Life Sciences, University of Nottingham, Nottingham NG7 2UH, UK; Centre of Membrane Proteins and Receptors, University of Birmingham and University of Nottingham, The Midlands, UK; Corresponding author
Erica K. Sloan
Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, VIC 3052, Australia; Cousins Center for Neuroimmunology, Semel Institute for Neuroscience and Human Behavior, Jonsson Comprehensive Cancer Center, and UCLA AIDS Institute, University of California Los Angeles, Los Angeles, CA 90095, USA; Division of Surgical Oncology, Peter MacCallum Cancer Centre, Victorian Comprehensive Cancer Centre, 305 Grattan Street, Melbourne, VIC 3000, Australia; Corresponding author
Summary: The therapeutic action of a drug depends on its ability to engage with its molecular target in vivo. However, current drug discovery strategies quantify drug levels within organs rather than determining the binding of drugs directly to their specific molecular targets in vivo. This is a particular problem for assessing the therapeutic potential of drugs that target malignant tumors where access and binding may be impaired by disrupted vasculature and local hypoxia. Here we have used triple-negative human breast cancer cells expressing β2-adrenoceptors tagged with the bioluminescence protein NanoLuc to provide a bioluminescence resonance energy transfer approach to directly quantify ligand binding to a G protein-coupled receptor in vivo using a mouse model of breast cancer. : Biological Sciences Tools; Cancer; Molecular Interaction; Optical Imaging Subject Areas: Biological Sciences Tools, Cancer, Molecular Interaction, Optical Imaging
