%0 Journal Article %1 Lohse1988 %A Lohse, M. J. %A Elger, B. %A Lindenborn-Fotinos, J. %A Klotz, K. N. %A Schwabe, U. %D 1988 %J Naunyn Schmiedebergs Arch Pharmacol %K & Acids Adenosine-5'-(N-ethylcarboxamide) Adenosine/*analogs Assay Binding Blood Cholic Chromatography, Gel Humans Platelets/*metabolism Radioligand Sites Solubility alpha/*isolation derivatives/metabolism purification Receptor Adrenergic %N 1 %P 64-8 %T Separation of solubilized A2 adenosine receptors of human platelets from non-receptor 3HNECA binding sites by gel filtration %U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2835689 %V 337 %X 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 3HNECA (5'-N-ethylcarboxamidoadenosine) was measured to the eluted fractions. Two 3HNECA binding peaks were eluted, the first of them with the void volume. This first peak represented between 10% and 25% of the 3HNECA binding activity eluted from the column. It bound 3HNECA 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 3HNECA 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 3HNECA 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 3HNECA binding sites by gel filtration. This allows reliable radioligand binding studies of the A2 adenosine receptor of human platelets.

@article{Lohse1988, 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.}, added-at = {2010-12-14T18:12:02.000+0100}, author = {Lohse, M. J. and Elger, B. and Lindenborn-Fotinos, J. and Klotz, K. N. and Schwabe, U.}, biburl = {https://www.bibsonomy.org/bibtex/2494cb4a967410d23d677de7c1a08be3f/pharmawuerz}, endnotereftype = {Journal Article}, interhash = {ddb5f7eec1fe210e7827c09117c0bb3d}, intrahash = {494cb4a967410d23d677de7c1a08be3f}, issn = {0028-1298 (Print) 0028-1298 (Linking)}, journal = {Naunyn Schmiedebergs Arch Pharmacol}, keywords = {& Acids Adenosine-5'-(N-ethylcarboxamide) Adenosine/*analogs Assay Binding Blood Cholic Chromatography, Gel Humans Platelets/*metabolism Radioligand Sites Solubility alpha/*isolation derivatives/metabolism purification Receptor Adrenergic}, month = Jan, note = {Lohse, M J Elger, B Lindenborn-Fotinos, J Klotz, K N Schwabe, U Germany, west Naunyn-Schmiedeberg's archives of pharmacology Naunyn Schmiedebergs Arch Pharmacol. 1988 Jan;337(1):64-8.}, number = 1, pages = {64-8}, shorttitle = {Separation of solubilized A2 adenosine receptors of human platelets from non-receptor [3H]NECA binding sites by gel filtration}, timestamp = {2010-12-14T18:22:41.000+0100}, title = {Separation of solubilized A2 adenosine receptors of human platelets from non-receptor [3H]NECA binding sites by gel filtration}, url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=2835689}, volume = 337, year = 1988 }