Partial separation of platelet and placental adenosine receptors from adenosine A2-like binding protein.
The ubiquitous adenosine A2-like binding protein obscures the binding properties of adenosine receptors assayed with 5'-N-[3H]ethylcarboxamidoadenosine [( 3H]NECA). To solve this problem, we developed a rapid and simple method to separate adenosine receptors from the adenosine A2-like binding protein. Human platelet and placental membranes were solubilized with 1% 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate. The soluble platelet extract was precipitated with polyethylene glycol and the fraction enriched in adenosine receptors was isolated from the precipitate by differential centrifugation. The adenosine A2-like binding protein was removed from the soluble placental extract with hydroxylapatite and adenosine receptors were precipitated with polyethylene glycol. The specificity of the [3H]NECA binding is typical of an adenosine A2 receptor for platelets and an adenosine A1 receptor for placenta. This method leads to enrichment of adenosine A2 receptors for platelets and adenosine A1 receptors for placenta. This provides a useful preparation technique for pharmacologic studies of adenosine receptors.
Zolnierowicz
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Department of Internal Medicine and Biological Chemistry, University Hospital, Ann Arbor, Michigan 48109-0108.
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Hutchison
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Fox
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D000241:Adenosine / Q000031:analogs & derivatives / Q000494:pharmacology; D019830:Adenosine-5'-(N-ethylcarboxamide); D001665:Binding Sites; D001792:Blood Platelets / Q000032:analysis*; D002352:Carrier Proteins / Q000302:isolation & purification*; D002462:Cell Membrane / Q000032:analysis; D006801:Humans; D010920:Placenta / Q000032:analysis*; D011869:Radioligand Assay; D011983:Receptors, Purinergic / Q000187:drug effects / Q000302:isolation & purification* / Q000378:metabolism; D014316:Tritium; D014665:Vasodilator Agents / Q000494:pharmacology
D002352:Carrier Proteins; D011983:Receptors, Purinergic; D014665:Vasodilator Agents; D014316:Tritium; D019830:Adenosine-5'-(N-ethylcarboxamide); D000241:Adenosine