| pubmed-article:3010272 | rdf:type | pubmed:Citation | lld:pubmed |
| pubmed-article:3010272 | lifeskim:mentions | umls-concept:C0010453 | lld:lifeskim |
| pubmed-article:3010272 | lifeskim:mentions | umls-concept:C1512977 | lld:lifeskim |
| pubmed-article:3010272 | lifeskim:mentions | umls-concept:C0034785 | lld:lifeskim |
| pubmed-article:3010272 | lifeskim:mentions | umls-concept:C1998793 | lld:lifeskim |
| pubmed-article:3010272 | pubmed:issue | 5 | lld:pubmed |
| pubmed-article:3010272 | pubmed:dateCreated | 1986-6-2 | lld:pubmed |
| pubmed-article:3010272 | pubmed:abstractText | S49 Mouse lymphoma wild-type cells were grown in spinner cultures of 40 liters to a density of approximately 3 million cells/ml. Growth of cells to high density (2-3 million cells/ml) required that the cell suspensions be bubbled with oxygen. Cells from 40 liter cultures were collected by centrifugation and disrupted by nitrogen cavitation. Highly purified membranes (0.35 g membrane protein) that were rich in beta-adrenergic receptor (0.4-0.7 pmol receptor/mg membrane protein) were prepared by differential centrifugation and then solubilized with the plant glycoside, digitonin (1.5% digitonin at 3 mg of membrane protein/ml). Beta-adrenergic receptors were isolated and purified by sequential affinity chromatography, ion-exchange chromatography, and steric exclusion high-pressure liquid chromatography. The extract was subjected to affinity chromatography on a derivatized Sepharose-4B CL column to which the high-affinity, beta-adrenergic antagonist (-)alprenolol had been immobilized. Following extensive washing, the receptor bound to this matrix was eluted using a 0-100 micromolar linear gradient of (-)alprenolol. The receptor eluted as a sharp peak at 30 micromolar ligand and displayed a specific activity of 280 pmol receptor/mg of protein. Ion-exchange chromatography on DEAE-Sephacel increased the specific activity to 950 pmol/mg of protein. The final step in the purification, steric-exclusion high-pressure liquid chromatography on two TSK-3000 and one TSK-2000 columns, tandem linked, resulted in a beta-adrenergic receptor preparation with a specific activity of 6700 pmol/mg of protein (15,900-fold purification). Autoradiography of the radioiodinated pure receptor, the receptor photolabeled with [125I]iodoazidobenzylpindolol or silver-staining of chemical amounts of protein revealed that the Mr of the pure receptor is 66,000 upon polyacrylamide gel electrophoresis in sodium dodecyl sulfate under reducing conditions. The receptor is a beta2-subtype adrenergic receptor. | lld:pubmed |
| pubmed-article:3010272 | pubmed:grant | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:3010272 | pubmed:grant | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:3010272 | pubmed:grant | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:3010272 | pubmed:language | eng | lld:pubmed |
| pubmed-article:3010272 | pubmed:journal | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:3010272 | pubmed:citationSubset | IM | lld:pubmed |
| pubmed-article:3010272 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:3010272 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:3010272 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:3010272 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:3010272 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:3010272 | pubmed:issn | 0032-7484 | lld:pubmed |
| pubmed-article:3010272 | pubmed:author | pubmed-author:MalbonC CCC | lld:pubmed |
| pubmed-article:3010272 | pubmed:author | pubmed-author:GeorgeS TST | lld:pubmed |
| pubmed-article:3010272 | pubmed:issnType | Print | lld:pubmed |
| pubmed-article:3010272 | pubmed:volume | 15 | lld:pubmed |
| pubmed-article:3010272 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:3010272 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:3010272 | pubmed:pagination | 349-66 | lld:pubmed |
| pubmed-article:3010272 | pubmed:dateRevised | 2007-11-14 | lld:pubmed |
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| pubmed-article:3010272 | pubmed:meshHeading | pubmed-meshheading:3010272-... | lld:pubmed |
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| pubmed-article:3010272 | pubmed:meshHeading | pubmed-meshheading:3010272-... | lld:pubmed |
| pubmed-article:3010272 | pubmed:meshHeading | pubmed-meshheading:3010272-... | lld:pubmed |
| pubmed-article:3010272 | pubmed:year | 1985 | lld:pubmed |
| pubmed-article:3010272 | pubmed:articleTitle | Large-scale purification of beta-adrenergic receptors from mammalian cells in culture. | lld:pubmed |
| pubmed-article:3010272 | pubmed:publicationType | Journal Article | lld:pubmed |
| pubmed-article:3010272 | pubmed:publicationType | Research Support, U.S. Gov't, P.H.S. | lld:pubmed |
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| http://linkedlifedata.com/r... | pubmed:referesTo | pubmed-article:3010272 | lld:pubmed |
| http://linkedlifedata.com/r... | pubmed:referesTo | pubmed-article:3010272 | lld:pubmed |
