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Separation of solubilized A2 adenosine receptors of human platelets from non-receptor [3H]NECA binding sites by gel filtration

Lohse, Martin J. and Elger, Bernice and Lindenborn-Fotinos, Jutta and Klotz, Karl-Norbert and Schwabe, Ulrich. (1988) Separation of solubilized A2 adenosine receptors of human platelets from non-receptor [3H]NECA binding sites by gel filtration. Naunyn-Schmiedeberg's Archives of Pharmacology, 337 (1). pp. 64-68.

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Official URL: https://edoc.unibas.ch/75062/

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Abstract

Human platelet membranes were solubilized with the zwitterionic detergent CHAPS (3-[3-(cholamidopropyl)-dimethylammonio]-1-propanesulfonate) and the solubilized extract subjected to gel filtration. Binding of the adenosine receptor agonist [3H]NECA (5'-N-ethylcarboxamidoadenosine) was measured to the eluted fractions. Two [3H]NECA binding peaks were eluted, the first of them with the void volume. This first peak represented between 10% and 25% of the [3H]NECA binding activity eluted from the column. It bound [3H]NECA in a reversible, saturable and GTP-dependent manner with an affinity of 46 nmol/l and a binding capacity of 510 fmol/mg protein. Various adenosine receptor ligands competed for the binding of [3H]NECA to the first peak with a pharmacological profile characteristic for the A2 adenosine receptor as determined from adenylate cyclase experiments. In contrast, most adenosine receptor ligands did not compete for [3H]NECA binding to the second, major peak. These results suggest that a solubilized A2 receptor-Gs protein complex of human platelets can be separated from other [3H]NECA binding sites by gel filtration. This allows reliable radioligand binding studies of the A2 adenosine receptor of human platelets.
Faculties and Departments:03 Faculty of Medicine > Departement Public Health > Ehemalige Einheiten Public Health > Medizin- und Gesundheitsethik (Reiter-Theil)
08 Cross-disciplinary Subjects > Ethik > Institut für Bio- und Medizinethik > Bio- und Medizinethik (Elger)
03 Faculty of Medicine > Departement Public Health > Ethik in der Medizin > Bio- und Medizinethik (Elger)
UniBasel Contributors:Elger, Bernice Simone
Item Type:Article, refereed
Article Subtype:Research Article
Publisher:Springer
ISSN:0028-1298
e-ISSN:1432-1912
Note:Publication type according to Uni Basel Research Database: Journal article
Identification Number:
Last Modified:27 May 2020 12:28
Deposited On:27 May 2020 12:28

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