G Protein-Coupled Receptors in Drug Discovery

1. Front Matter

   Pages i-xi

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2. New Insights into GPCR Function: Implications for HTS

   • Richard M. Eglen, Terry Reisine

   Pages 1-13

3. Screening Technologies for G Protein-Coupled Receptors: From HTS to uHTS

   • Maite de los Frailes, Emilio Diez

   Pages 15-37

4. GPCR Signaling: Understanding the Pathway to Successful Drug Discovery

   • Christine Williams, Stephen J. Hill

   Pages 39-50

5. An Overview on GPCRs and Drug Discovery: Structure-Based Drug Design and Structural Biology on GPCRs

   • Kenneth Lundstrom

   Pages 51-66

6. Understanding the Ligand–Receptor–G Protein Ternary Complex for GPCR Drug Discovery

   • Venkata R.P. Ratnala, Brian Kobilka

   Pages 67-77

7. Assay Data Quality Assessment

   • Hanspeter Gubler

   Pages 79-95

8. Homology Modeling of GPCRs

   • John Simms, Nathan E. Hall, Polo H.C. Lam, Laurence J. Miller, Arthur Christopoulos, Ruben Abagyan et al.

   Pages 97-113

9. GPCR Expression Using Baculovirus-Infected Sf9 Cells

   • Amanda L. Aloia, Richard V. Glatz, Edward J. McMurchie, Wayne R. Leifert

   Pages 115-129

10. Radioligand Binding Assays: Application of [125I]Angiotensin II Receptor Binding

    • Wayne R. Leifert, Olgatina Bucco, Mahinda Y. Abeywardena, Glen S. Patten

    Pages 131-141

11. [35S]GTPγS Binding in G Protein-Coupled Receptor Assays

    • Tamara Cooper, Edward J. McMurchie, Wayne R. Leifert

    Pages 143-151

12. A Time-Resolved Fluorescent Lanthanide (Eu)-GTP Binding Assay for Chemokine Receptors as Targets in Drug Discovery

    • Jean Labrecque, Rebecca S.Y. Wong, Simon P. Fricker

    Pages 153-169

13. Use of the DiscoveRx Hithunter cAMPII Assay for Direct Measurement of cAMP in Gs and Gi GPCRs

    • Joe Bradley, David McLoughlin

    Pages 171-179

14. Use of Aequorin for G protein-Coupled Receptor Hit Identification and Compound Profiling

    • Stephen J. Brough, Parita Shah

    Pages 181-198

15. BacMam: Versatile Gene Delivery Technology for GPCR Assays

    • Elizabeth A. Davenport, Parvathi Nuthulaganti, Robert S. Ames

    Pages 199-211

16. Yeast Assays for G Protein-Coupled Receptors

    • Simon J. Dowell, Andrew J. Brown

    Pages 213-229

17. GPCR Microspot Assays on Solid Substrates

    • Ye Fang, Joydeep Lahiri

    Pages 231-238

18. Resonant Waveguide Grating Biosensor for Whole-Cell GPCR Assays

    • Ye Fang, Ann M. Ferrie, Elizabeth Tran

    Pages 239-252

19. FRET-Based Measurement of GPCR Conformational Changes

    • Sébastien Granier, Samuel Kim, Juan José Fung, Michael P. Bokoch, Charles Parnot

    Pages 253-268

20. FLIPR® Assays of Intracellular Calcium in GPCR Drug Discovery

    • Kasper B. Hansen, Hans Bräuner-Osborne

    Pages 269-278

The G protein-coupled receptors (GPCRs) and associated peripheral G proteins underpin a multitude of physiological processes. The GPCRs represent one of the largest superfamilies in the human genome and are a significant target for bioactive and drug discovery programs. It is estimated that greater than 50% of all drugs, including those in development, currently target GPCRs. Many of the characterized GPCRs have known ligands; however, approximately 20% of GPCRs are described as orphan GPCRs, apparent GPCRs that share the generic high-level structure charact- istic of GPCRs but whose endogenous ligand is not known. Therefore, it is expected that the field of GPCR drug discovery and development will greatly expand in the coming years with emphasis on new generations of drugs against GPCRs with unique therapeuticuseswhichmayincludedrugssuchasallostericregulators,inverseagonists, and identification of orphan GPCR ligands. AswelearnmoreaboutthemolecularsignalingcascadesfollowingGPCRactivation, we acquire a better appreciation of the complexity of cell signaling and as a result, also acquire a vast array ofnew molecularmethods toinvestigate these andother processes. Thegeneralaimofthisbookistoprovideresearcherswitharangeofprotocolsthatmay be useful in their GPCR drug discovery programs. It is also the basis for the devel- ment of future assays in this field. Therefore, the range of topics covered and the appropriate methodological approaches in GPCR drug discovery are reflected in this book. Itisinterestingtonotethatfuturedirectionsindrugdiscoverywillrequireinput and collaboration from a plethora of fields of research. As such, this book will likely be of interest to scientists involved in such fields as molecular biology, pharmacology, biochemistry, cellular signaling, andbio-nanotechnology.

• Calcium
• Cellular signalling
• Chemokine
• Drug development
• GPCRs
• Ligands
• physiology

From the reviews: “To fully understand the GPCR drug discovery process one should … have wide knowledge of the methods/protocols used in this process. This book serves this exact purpose. … It’s biggest strength is a clear description of every aspect of GPCR drug discovery process, which is easy to understand even for a non-expert in the respective field. … a must-read for everyone involved in GPCR drug discovery and particularly for scientists working in multidisciplinary teams, who need to soundly communicate ideas with experts in other fields.” (Bartosz Trzaskowski, Acta Biochimica Polonica, Vol. 56, December, 2009)

• CSIRO Human Nutrition, Adelaide, Australia

  Wayne R. Leifert

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