Abstract
A wide range of peptides and polypeptides can be appended to either the N- or C-terminus of G proteincoupled receptors without disrupting substantially ligand binding and signal transduction. Following fusion of fluorescent proteins, reporter gene constructs or G protein α subunits to the C-terminal tail of a receptor high content and G protein activation assays can be employed to identify agonist ligands. Further modification of the receptor fusions to introduce enhanced levels of constitutive activity and to physically destabilise the protein allows antagonist / inverse agonists screens to be developed in parallel. Equivalent C-terminal addition of pairs of complementary, non-functional, polypeptide fragments allows the application of enzyme complementation techniques. Introduction of N-terminal tags to receptors has also allowed the introduction of novel assay techniques based on a pH-sensitive cyanine dye. These have the capacity to overcome certain limitations of GPCR-fluorescent protein fusions.
Keywords: assay development, g protein-coupled receptor, g protein, enzyme complementation, green fluorescent protein
Current Pharmaceutical Design
Title: G Protein-Coupled Receptor Fusion Proteins in Drug Discovery
Volume: 10 Issue: 17
Author(s): G. Milligan, G- J. Feng, R. J. Ward, N. Sartania, D. Ramsay, A. J. McLean and J. J. Carrillo
Affiliation:
Keywords: assay development, g protein-coupled receptor, g protein, enzyme complementation, green fluorescent protein
Abstract: A wide range of peptides and polypeptides can be appended to either the N- or C-terminus of G proteincoupled receptors without disrupting substantially ligand binding and signal transduction. Following fusion of fluorescent proteins, reporter gene constructs or G protein α subunits to the C-terminal tail of a receptor high content and G protein activation assays can be employed to identify agonist ligands. Further modification of the receptor fusions to introduce enhanced levels of constitutive activity and to physically destabilise the protein allows antagonist / inverse agonists screens to be developed in parallel. Equivalent C-terminal addition of pairs of complementary, non-functional, polypeptide fragments allows the application of enzyme complementation techniques. Introduction of N-terminal tags to receptors has also allowed the introduction of novel assay techniques based on a pH-sensitive cyanine dye. These have the capacity to overcome certain limitations of GPCR-fluorescent protein fusions.
Export Options
About this article
Cite this article as:
Milligan G., Feng J. G-, Ward J. R., Sartania N., Ramsay D., McLean J. A. and Carrillo J. J., G Protein-Coupled Receptor Fusion Proteins in Drug Discovery, Current Pharmaceutical Design 2004; 10 (17) . https://dx.doi.org/10.2174/1381612043384295
DOI https://dx.doi.org/10.2174/1381612043384295 |
Print ISSN 1381-6128 |
Publisher Name Bentham Science Publisher |
Online ISSN 1873-4286 |
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
- Announcements
Related Articles
-
Withdrawal Notice: Drug Repurposing for Prospective Anti-Cancer Agents Along with the Clinical Status of the Repurposed Drug
Anti-Cancer Agents in Medicinal Chemistry Molecular Targeting of Cell Death Signal Transduction Pathways in Cancer
Current Signal Transduction Therapy Left-Right Asymmetry in Embryonic Development and Breast Cancer: Common Molecular Determinants?
Current Medicinal Chemistry An Update on Overcoming MDR1-Mediated Multidrug Resistance in Cancer Chemotherapy
Current Pharmaceutical Design Subdural Pharmacotherapy for the Treatment of Intractable Focal Neocortical Epilepsy
Drug Delivery Letters Recent Advances in Optical Cancer Imaging of EGF Receptors
Current Medicinal Chemistry Xanthones from Mangosteen Extracts as Natural Chemopreventive Agents: Potential Anticancer Drugs
Current Molecular Medicine Multivariate Statistical Tools for the Evaluation of Proteomic 2D-maps:Recent Achievements and Applications
Current Proteomics Gliomas: New Perspectives in Diagnosis, Treatment and Prognosis
Current Topics in Medicinal Chemistry Anti-EGFR Therapy: Strategies in Head and Neck Squamous Cell Carcinoma
Recent Patents on Anti-Cancer Drug Discovery Microglia Phenotype Diversity
CNS & Neurological Disorders - Drug Targets Patent Selections:
Current Biomarkers (Discontinued) Specific Cytostatic and Cytotoxic Effect of Dihydrochelerythrine in Glioblastoma Cells: Role of NF-κB/β-catenin and STAT3/IL-6 Pathways
Anti-Cancer Agents in Medicinal Chemistry Targeting SREBP-1-driven Lipid Metabolism to Treat Cancer
Current Pharmaceutical Design Gene Expression Analysis Approach to Establish Possible Links Between Parkinson's Disease, Cancer and Cardiovascular Diseases
CNS & Neurological Disorders - Drug Targets The Urokinase Receptor as a Potential Target in Cancer Therapy
Current Pharmaceutical Design Stem Cell and Regenerative Medicine
Current Stem Cell Research & Therapy Biochemical Markers for Brain Injury Monitoring in Children with or without Congenital Heart Diseases
CNS & Neurological Disorders - Drug Targets Ion Transporters in Brain Tumors
Current Medicinal Chemistry β-Catenin/TCF-4 Signaling Regulates Susceptibility of Macrophages and Resistance of Monocytes to HIV-1 Productive Infection
Current HIV Research