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Review

GTPγS Binding Assays

Neil W. DeLapp  1 , Wendy H. Gough, Steven D. Kahl, Amy C. Porter, Todd R. Wiernicki  1
Sarine Markossian  1 , Abigail Grossman  1 , Hannah Baskir  1 , Michelle Arkin  2 , Douglas Auld  3 , Christopher Austin  4 , Jonathan Baell  5 , Kyle Brimacombe  1 , Thomas D.Y. Chung  6 , Nathan P. Coussens  7 , Jayme L. Dahlin  8 , Viswanath Devanarayan  9 , Timothy L. Foley  10 , Marcie Glicksman  11 , Kirill Gorshkov  12 , Stefan Grotegut  13 , Matthew D. Hall  1 , Samuel Hoare  14 , James Inglese  1 , Philip W. Iversen  15 , Madhu Lal-Nag  16 , Zhuyin Li  12 , Jason R. Manro  13 , James McGee  17 , Allison Norvil  13 , Mackenzie Pearson  13 , Terry Riss  18 , Peter Saradjian  19 , G. Sitta Sittampalam  20 , Michael A. Tarselli  21 , O. Joseph Trask Jr.  22 , Jeffrey R. Weidner  23 , Mary Jo Wildey  24 , Kelli Wilson  1 , Menghang Xia  1 , Xin Xu  1
, editors.
In: Assay Guidance Manual [Internet]. Bethesda (MD): Eli Lilly & Company and the National Center for Advancing Translational Sciences; 2004.
[updated ].
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Review

GTPγS Binding Assays

Neil W. DeLapp et al.
Free Books & Documents

Excerpt

Signal transduction via G-protein coupled receptors (GPCRs) is mediated through binding of GTP to GTP‑binding proteins. Evaluation of GPCR agonists in drug discovery requires GTP binding assays to confirm agonist activity. This chapter addresses the development and optimization of robust GTPγS binding assays with discussions on basic protocols, assay buffers, effect of required additives (Mg2+, NaCl, Saponin) and data analysis. An extensive set of recent references is included for beginners and experienced investigators.

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References

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Additional References

    1. Cussac D, Newman-Tancredi A, Duqueyroix D, Pasteau V, Millan MJ. Differential activation of Gq/11 and Gi3 proteins at 5-HT2C receptors revealed by antibody capture assays: influence of receptor reserve and relationship to agonist-directed trafficking. Mol. Pharmacol. 2002;62:578–58. - PubMed
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    1. Eglen RM, Reisine T. New Insights into GPCR Function: Implications for HTS. In: G-Protein Coupled Receptors in Drug Discovery, series. Methods in Molecular Biology. 2009;551:1–13. - PubMed

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