Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
12
|
| pubmed:dateCreated |
1993-4-27
|
| pubmed:abstractText |
An expression plasmid encoding the extracellular domain of the 75-kDa human tumor necrosis factor (TNF) type 2 receptor (TNF-R2) was constructed and used to generate a stable cell line secreting soluble TNF-R2 (sTNF-R2). Purified sTNF-R2 was resolved by SDS-PAGE into one band of approximate M(r) 43,000, consistent with a molecular weight of 36,000 +/- 4800 obtained by sedimentation equilibrium analysis. The apparent molecular weight observed by gel filtration chromatography was approximately 136,000. Glycosylation analysis revealed that Asn-149 is fully glycosylated, while Asn-171 is incompletely glycosylated (approximately 50%), and that a proline-, serine-, and threonine-rich region (residues 175-234) contains O-linked carbohydrate structures. Scatchard analysis of [125I]TNF-alpha and [125I]TNF-beta binding to sTNF-R2 gave dissociation constants (Kd) of 0.3 and 0.75 nM, respectively, comparable to those observed for intact cell-surface TNF-R2. The sTNF-R2 was found to block the cytotoxicity of both TNF-alpha and TNF-beta in a murine L-M cell assay. The sizes of the sTNF-R2.TNF-alpha and sTNF-R2.TNF-beta complexes determined by gel filtration chromatography were approximately 322 and 204 kDa, respectively. The stoichiometry of the sTNF-R2.TNF-alpha and sTNF-R2.TNF-beta complexes were examined by size-exclusion chromatography, sedimentation equilibrium, and cross-linking. The data from these studies suggest that at least two molecules of sTNF-R2 can bind to a single TNF-alpha or TNF-beta trimer.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Cross-Linking Reagents,
http://linkedlifedata.com/resource/pubmed/chemical/Lymphotoxin-alpha,
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/Tumor Necrosis Factor-alpha
|
| pubmed:status |
MEDLINE
|
| pubmed:month |
Mar
|
| pubmed:issn |
0006-2960
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
30
|
| pubmed:volume |
32
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
3131-8
|
| pubmed:dateRevised |
2007-11-15
|
| pubmed:meshHeading |
pubmed-meshheading:8384489-Amino Acid Sequence,
pubmed-meshheading:8384489-Animals,
pubmed-meshheading:8384489-Carbohydrate Conformation,
pubmed-meshheading:8384489-Cell Line,
pubmed-meshheading:8384489-Chromatography, Gel,
pubmed-meshheading:8384489-Chromatography, High Pressure Liquid,
pubmed-meshheading:8384489-Cross-Linking Reagents,
pubmed-meshheading:8384489-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:8384489-Embryo, Mammalian,
pubmed-meshheading:8384489-Glycosylation,
pubmed-meshheading:8384489-Humans,
pubmed-meshheading:8384489-Kidney,
pubmed-meshheading:8384489-Lymphotoxin-alpha,
pubmed-meshheading:8384489-Mice,
pubmed-meshheading:8384489-Molecular Sequence Data,
pubmed-meshheading:8384489-Molecular Weight,
pubmed-meshheading:8384489-Receptors, Cell Surface,
pubmed-meshheading:8384489-Receptors, Tumor Necrosis Factor,
pubmed-meshheading:8384489-Transfection,
pubmed-meshheading:8384489-Tumor Necrosis Factor-alpha,
pubmed-meshheading:8384489-Ultracentrifugation
|
| pubmed:year |
1993
|
| pubmed:articleTitle |
Biochemical characterization of the extracellular domain of the 75-kilodalton tumor necrosis factor receptor.
|
| pubmed:affiliation |
Department of Molecular Biology, Genentech, Inc., South San Francisco, California 94080.
|
| pubmed:publicationType |
Journal Article
|