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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
33
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pubmed:dateCreated |
1990-12-28
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pubmed:abstractText |
Two distinct tumor necrosis factor (TNF) receptors of 55- and 75-kDa apparent molecular masses previously identified on the cell surface by monoclonal antibodies have been solubilized with Triton X-100 from HL60 cells. A filter-based dot blot assay was developed to monitor specific 125I-TNF alpha binding during fractionation of the cell extract. By a combination of immuno- and ligand affinity chromatography and reverse phase high performance liquid chromatography both receptor proteins were purified to apparent homogeneity. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed two bands at 55 and 51 kDa for the 55-kDa TNF receptor and a major 75-kDa and a minor 65-kDa band for the 75-kDa TNF receptor. All these bands specifically bound TNF alpha and TNF beta in ligand blot experiments. The exclusive specificity of monoclonal antibodies of the utr series for the 75.65-kDa bands and of the htr series for the 55.51-kDa bands was demonstrated with the purified antigens on Western blots. Both TNF receptor types were found to contain N-linked carbohydrates. N-terminal amino acid sequence analysis of the 55- and 51-kDa bands of the 55-kDa TNF receptor revealed identical sequences suggesting a possible truncation at the C-terminal end. Two different N-terminal sequences were determined for the 65-kDa band. One corresponded to the published sequence of ubiquitin; the other was therefore assumed to be a unique sequence of the 75-kDa TNF receptor. Additional internal sequences of this receptor were determined after proteolytic cleavage.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Peptide Fragments,
http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Cell Surface,
http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Tumor Necrosis Factor,
http://linkedlifedata.com/resource/pubmed/chemical/Trypsin,
http://linkedlifedata.com/resource/pubmed/chemical/Tumor Necrosis Factor-alpha
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pubmed:status |
MEDLINE
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pubmed:month |
Nov
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pubmed:issn |
0021-9258
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
25
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pubmed:volume |
265
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
20131-8
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pubmed:dateRevised |
2004-11-17
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pubmed:meshHeading |
pubmed-meshheading:2173696-Amino Acid Sequence,
pubmed-meshheading:2173696-Cell Line,
pubmed-meshheading:2173696-Chromatography, Affinity,
pubmed-meshheading:2173696-Chromatography, High Pressure Liquid,
pubmed-meshheading:2173696-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:2173696-Humans,
pubmed-meshheading:2173696-Kinetics,
pubmed-meshheading:2173696-Leukemia, Promyelocytic, Acute,
pubmed-meshheading:2173696-Molecular Sequence Data,
pubmed-meshheading:2173696-Molecular Weight,
pubmed-meshheading:2173696-Peptide Fragments,
pubmed-meshheading:2173696-Receptors, Cell Surface,
pubmed-meshheading:2173696-Receptors, Tumor Necrosis Factor,
pubmed-meshheading:2173696-Trypsin,
pubmed-meshheading:2173696-Tumor Necrosis Factor-alpha
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pubmed:year |
1990
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pubmed:articleTitle |
Purification and partial amino acid sequence analysis of two distinct tumor necrosis factor receptors from HL60 cells.
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pubmed:affiliation |
Central Research Units, F. Hoffmann-La Roche Ltd., Basel, Switzerland.
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pubmed:publicationType |
Journal Article
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