[Analysis of the TNF receptor in KYM cells by affinity cross-linking].
We investigated the identity of the TNF receptor on the KYM cell membrane by cross-linking 125I-TNF and the presumed receptor site with DSS, and subjecting the TNF-receptor complex to electrophoresis. Four specific bands were observed at 145 K, 50 K, 35 K, and 17 K, and those at 50 K, 35 K and 17 K being consistent with trimers, dimers and monomers of TNF, respectively. The 145 K band disappeared after addition of excess unlabelled TNF or anti-human recombinant TNF monoclonal antibody (IV3-E), which quenched the cytotoxic activity of TNF and inhibited the TNF binding to the receptor. The molecular weight of native TNF as estimated by gel filtration was 45 K and this observation showed that native TNF existed only as the TNF trimer. These results confirmed that 95K, i.e., the difference between 145 K and 50 K, is the molecular size of the TNF receptor.
D000911:Antibodies, Monoclonal / Q000276:immunology; D002462:Cell Membrane / Q000032:analysis; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003432:Cross-Linking Reagents; D006023:Glycoproteins / Q000276:immunology / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis; D008970:Molecular Weight; D009217:Myosarcoma / Q000032:analysis* / Q000473:pathology; D011956:Receptors, Cell Surface / Q000032:analysis*; D018124:Receptors, Tumor Necrosis Factor; D012509:Sarcoma / Q000032:analysis*; D014409:Tumor Necrosis Factor-alpha
D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D006023:Glycoproteins; D008565:Membrane Proteins; D011956:Receptors, Cell Surface; D018124:Receptors, Tumor Necrosis Factor; D014409:Tumor Necrosis Factor-alpha