Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
35
|
| pubmed:dateCreated |
1989-1-20
|
| pubmed:abstractText |
The receptor for human tumor necrosis factor-alpha (TNF-alpha) was isolated from a subclone of the human histiocytic lymphoma cell line U937. These cells exhibit a single class of high affinity receptors (Kd = 0.51 +/- 0.25 nM) with an average density of 55,000 +/- 5,000 binding sites/cell. After solubilization with detergent, the receptor retained its ability to bind free TNF-alpha but failed to bind to TNF-alpha immobilized on various solid supports. For receptor purification, 125I-TNF-alpha was covalently attached to the receptor on intact cells by the bifunctional cross-linking reagents ethylene glycolbis(succinimidylsuccinate) or 3,3-dithiobis(sulfosuccinimidylpropionate). The cells were then solubilized with the nonionic detergent Triton X-100, and the supernatants, clarified by centrifugation, were passed over an IgG-Sepharose column prepared from TNF-alpha antiserum. The receptor-rich fraction from the antibody column was further purified by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These two steps together provided approximately 165,000-fold purification of the TNF-alpha receptor. The TNF-alpha receptor-ligand complex obtained by this method had a subunit molecular weight of 100,000 +/- 5,000 when examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis but on gel filtration the complex migrated with an apparent molecular weight of 480,000 +/- 32,000. However, the receptor showed a molecular weight of 65,000 +/- 32,000 when gel filtration was performed in the absence of ligand. Additional characteristics of the receptor are discussed.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Dec
|
| pubmed:issn |
0021-9258
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
15
|
| pubmed:volume |
263
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
19098-104
|
| pubmed:dateRevised |
2004-11-17
|
| pubmed:meshHeading |
pubmed-meshheading:2848815-Binding, Competitive,
pubmed-meshheading:2848815-Cell Line,
pubmed-meshheading:2848815-Chromatography, Affinity,
pubmed-meshheading:2848815-Chromatography, Gel,
pubmed-meshheading:2848815-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:2848815-Humans,
pubmed-meshheading:2848815-Kinetics,
pubmed-meshheading:2848815-Molecular Weight,
pubmed-meshheading:2848815-Receptors, Cell Surface,
pubmed-meshheading:2848815-Receptors, Tumor Necrosis Factor,
pubmed-meshheading:2848815-Solubility
|
| pubmed:year |
1988
|
| pubmed:articleTitle |
Human tumor necrosis factor-alpha receptor. Purification by immunoaffinity chromatography and initial characterization.
|
| pubmed:affiliation |
Department of Protein Chemistry, Genentech, Inc., South San Francisco, California 94080.
|
| pubmed:publicationType |
Journal Article
|