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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
2
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pubmed:dateCreated |
1983-9-23
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pubmed:abstractText |
Acetylcholine receptor, isolated in Triton X-100 on a cobra alpha-neurotoxin affinity column was incorporated into unilamellar phospholipid vesicles by a detergent depletion method using Amberlite XAD-2. Vesicles of an average diameter of 25 nm were formed, as verified by freeze-fracture electron microscopy and gel filtration. 85 to 95% of the alpha-bungarotoxin binding sites of the reconstituted acetylcholine receptor were oriented towards the outside of the vesicles. In the reconstituted receptor one molecule of residual Triton X-100 per 2.5 alpha-bungarotoxin binding sites on the receptor molecule could be assessed. The reconstituted protein was not accessible to papain digestion, whereas the pure acetylcholine receptor, solubilized by Triton X-100 was split into smaller polypeptides under the same condition. Reconstituted acetylcholine receptor and receptor-rich membranes did not exhibit the same behavior as measured by use of a potentiometric dye. This is interpreted as an irreversible alteration of at least 95% of the receptors purified in the presence of Triton X-100. Furthermore, it could be shown that the fluorescence intensity changes induced by carbamylcholine in receptor-rich membranes did not reflect ion fluxes, but conformational changes of the protein or a displacement of the dye from the protein.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:issn |
0022-2631
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
74
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
75-84
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:6876149-Animals,
pubmed-meshheading:6876149-Chromatography, Affinity,
pubmed-meshheading:6876149-Cobra Venoms,
pubmed-meshheading:6876149-Electric Organ,
pubmed-meshheading:6876149-Freeze Fracturing,
pubmed-meshheading:6876149-Liposomes,
pubmed-meshheading:6876149-Microscopy, Electron,
pubmed-meshheading:6876149-Phosphatidylcholines,
pubmed-meshheading:6876149-Potentiometry,
pubmed-meshheading:6876149-Receptors, Cholinergic,
pubmed-meshheading:6876149-Spectrometry, Fluorescence,
pubmed-meshheading:6876149-Torpedo
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pubmed:year |
1983
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pubmed:articleTitle |
Reconstitution of pure acetylcholine receptor in phospholipid vesicles and comparison with receptor-rich membranes by the use of a potentiometric dye.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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