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Separation of solubilized A\(_2\) adenosine receptors of human platelets from non-receptor [\(^3\)H]NECA binding sites by gel filtration
Please always quote using this URN: urn:nbn:de:bvb:20-opus-60309
- Human platelet membranes were solubilized with the zwitterionic detergent CHAPS (3-[3-(cholamidopropyl)dimethylammonio]- 1-propanesulfonate) and the solubilized extract subjected to gel ftltration. Binding of the adenosine receptor agonist [\(^3\)H]NECA (5'-N-ethylcarboxamidoadenosine) was measured to the eluted fractions. Two [\(^3\)H]NECA binding peaks were eluted, the first of them with the void volume. This first peak represented between 10% and 25% of the [\(^3\)H]NECA binding activity eluted from the column. It bound [\(^3\)H]NECA in aHuman platelet membranes were solubilized with the zwitterionic detergent CHAPS (3-[3-(cholamidopropyl)dimethylammonio]- 1-propanesulfonate) and the solubilized extract subjected to gel ftltration. Binding of the adenosine receptor agonist [\(^3\)H]NECA (5'-N-ethylcarboxamidoadenosine) was measured to the eluted fractions. Two [\(^3\)H]NECA binding peaks were eluted, the first of them with the void volume. This first peak represented between 10% and 25% of the [\(^3\)H]NECA binding activity eluted from the column. It bound [\(^3\)H]NECA in a reversible, saturable and GTPdependent manner with an affinity of 46 nmol/1 and a binding capacity of 510 fmol/mg protein. Various adenosine receptor ligands competed for the binding of [\(^3\)H]NECA to the frrst peak with a pharmacological proftle characteristic for the A\(_2\) adenosine receptor as determined from adenylate cyclase experiments. In contrast, most adenosine receptor ligands did not compete for [\(^3\)H]NECA binding to the second, major peak. These results suggest that a solubilized A\(_2\) receptor-Gs protein complex of human platelets can be separated from other [\(^3\)H]NECA binding sites by gel filtration. This allows reliable radioligand binding studies of the A2 adenosine receptor of human plate1ets.…
Author: | M. J. Lohse, B. Elger, J. Lindenborn-Fotinos, Karl-Norbert Klotz, U. Schwabe |
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URN: | urn:nbn:de:bvb:20-opus-60309 |
Document Type: | Journal article |
Faculties: | Medizinische Fakultät / Institut für Pharmakologie und Toxikologie |
Language: | English |
Year of Completion: | 1988 |
Source: | Naunyn-Schmiedeberg's Archives of Pharmacology (1988), 337, 64-68. |
Dewey Decimal Classification: | 6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit |
GND Keyword: | Toxikologie |
Tag: | A<sub>2</sub> Adenosine receptor; Adenylate cyclase; Human platelets; Radioligand binding |
Release Date: | 2011/12/08 |
Licence (German): | Deutsches Urheberrecht |