Tritium-labeled agonists as tools for studying adenosine A2B receptors

Purinergic Signalling
Sonja HinzChrista E Müller

Abstract

A selective agonist radioligand for A2B adenosine receptors (A2BARs) is currently not available. Such a tool would be useful for labeling the active conformation of the receptors. Therefore, we prepared BAY 60-6583, a potent and functionally selective A2BAR (partial) agonist, in a tritium-labeled form. Despite extensive efforts, however, we have not been able to establish a radioligand binding assay using [3H]BAY 60-6583. This is probably due to its high non-specific binding and its moderate affinity, which had previously been overestimated based on functional data. As an alternative, we evaluated the non-selective A2BAR agonist [3H]NECA for its potential to label A2BARs. [3H]NECA showed specific, saturable, and reversible binding to membrane preparations of Chinese hamster ovary (CHO) or human embryonic kidney (HEK) cells stably expressing human, rat, or mouse A2BARs. In competition binding experiments, the AR agonists 2-chloroadenosine (CADO) and NECA displayed significantly higher affinity when tested versus [3H]NECA than versus the A2B-antagonist radioligand [3H]PSB-603 while structurally diverse AR antagonists showed the opposite effects. Although BAY 60-6583 is an A2BAR agonist, it displayed higher affinity versus [3H]PSB...Continue Reading

References

Dec 1, 1989·Naunyn-Schmiedeberg's Archives of Pharmacology·K N KlotzM Grifantini
Jun 1, 1997·British Journal of Pharmacology·S DionisottiB B Fredholm
Dec 24, 1997·British Journal of Pharmacology·M MirabetR Franco
Jun 6, 2000·European Journal of Pharmaceutical Sciences : Official Journal of the European Federation for Pharmaceutical Sciences·C E MüllerR Sauer
Mar 27, 2001·Biochemical Pharmacology·X JiK A Jacobson
Jun 5, 2004·Bioorganic & Medicinal Chemistry Letters·Pier Giovanni BaraldiPier Andrea Borea
Jun 15, 2004·Biochemical Pharmacology·Mike StewartStephen J MacLennan
Mar 7, 2006·Nature Reviews. Drug Discovery·Kenneth A Jacobson, Zhan-Guo Gao
Mar 14, 2007·Circulation·Tobias EckleHolger K Eltzschig
Feb 3, 2009·Purinergic Signalling·Pier Giovanni BaraldiDelia Preti
Nov 6, 2009·Current Opinion in Pulmonary Medicine·Nicola A HananiaRichard A Bond
Dec 28, 2010·Biochimica Et Biophysica Acta·Christa E Müller, Kenneth A Jacobson
Mar 12, 2011·Science·Fei XuRaymond C Stevens
Oct 6, 2011·The Journal of Physical Chemistry. B·Yuli LiuEsteban Vöhringer-Martinez
Mar 19, 2014·The Journal of Pharmacology and Experimental Therapeutics·Sonja HinzChrista E Müller
Apr 1, 2014·Current Opinion in Cell Biology·James W WislerRobert J Lefkowitz
May 24, 2014·Biochemical Pharmacology·Zhan-Guo GaoKenneth A Jacobson
Mar 13, 2015·Molecular Pharmacology·Guillaume LebonChristopher G Tate
Jan 23, 2016·The Journal of Pharmacology and Experimental Therapeutics·Elizabeth A VecchioLauren T May
Jan 30, 2016·Journal of Medicinal Chemistry·Abdelaziz El MaatouguiEddy Sotelo
Apr 14, 2016·Current Topics in Medicinal Chemistry·Daniel T Gewirth
Jul 28, 2016·Nature Communications·Harald HübnerPeter Gmeiner
Nov 16, 2016·European Journal of Medicinal Chemistry·Sujay BasuKasim A Mookhtiar
Jan 28, 2017·Cell·Daniel WackerBryan L Roth
Feb 9, 2017·Proceedings of the National Academy of Sciences of the United States of America·Bingfa SunBrian K Kobilka
Feb 9, 2017·Immunology and Cell Biology·David AllardJohn Stagg
Feb 25, 2017·Cell·Alisa GlukhovaArthur Christopoulos

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Citations

Jan 25, 2019·Annual Review of Immunology·Joel LindenGerhard Dahl
Jan 30, 2019·Expert Opinion on Therapeutic Targets·Peter IllesYong Tang
Oct 20, 2019·International Journal of Molecular Sciences·Zhan-Guo Gao, Kenneth A Jacobson
Nov 16, 2019·Pharmaceuticals·Stephanie FedericoGiampiero Spalluto
Feb 14, 2021·Cellular and Molecular Life Sciences : CMLS·Rafael FrancoLeonardo Pardo
Dec 15, 2020·Purinergic Signalling·Xue YangDaan van der Es

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Methods Mentioned

BETA
competition binding
X-ray
transfection
saturation binding

Software Mentioned

Graph Pad Prism

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