PMID: 8947920Nov 1, 1996Paper

35S]GTP gamma S binding: a tool to evaluate functional activity of a cloned opioid receptor transiently expressed in COS cells

Neurochemical Research
K BefortB Kieffer

Abstract

In this paper we propose a powerful procedure to measure functional activation of the mouse delta-opioid receptor transiently expressed in mammalian cells. Receptor stimulation was assessed using a population of electroporated COS cells, transfected at a 50% efficiency. Under those conditions, agonist-promoted activation of the receptor was measured by [35S]GTP gamma S binding. Both BW373U86, an alkaloid compound, and DADLE, a peptide agonist, elicited increase of specific [35S]GTP gamma S binding representing 300% of basal level. Maximal activation was compared to that obtained for the cloned receptor stably expressed in CHO cells. Agonist efficacy was similar in both expressions systems, demonstrating the high sensitivity of the proposed method applied to transient expression. Finally dose-response curves were found highly reproducible across transfection experiments, opening the possibility for a direct comparison of distinct recombinant receptor preparations. This method represents a powerful tool for the study of opioid signal transduction at the receptor level. It may also be extended to investigate signalling properties of other Gi/Go coupled receptors.

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Citations

Dec 1, 2000·Biochemical and Biophysical Research Communications·K OkadaY Shimohigashi
May 22, 2002·European Journal of Pharmacology·Dauren BiyashevSándor Benyhe
Dec 13, 2000·Journal of Molecular and Cellular Cardiology·C L NeilanS F Bolling
Mar 11, 2000·The European Journal of Neuroscience·K MonoryA Borsodi
Feb 27, 2004·Protein Engineering, Design & Selection : PEDS·David OttAndreas Plückthun
Apr 25, 2003·Biopolymers·Georg SchlechtingenMurray Goodman
Apr 17, 2004·Journal of Neurochemistry·Frederic SimoninBrigitte L Kieffer
Oct 16, 2015·Anesthesia and Analgesia·Eszter SzlaviczEngin Bojnik
Jun 22, 1999·The Journal of Biological Chemistry·K BefortB L Kieffer
Apr 23, 1999·The Journal of Biological Chemistry·D A MasonS B Liggett
Dec 6, 1997·Biochemical and Biophysical Research Communications·M HiraiwaJ S O'Brien

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