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Molecular cloning and expression of human and rat tumor necrosis factor receptor chain (p60) and its soluble derivative, tumor necrosis factor-binding protein.
- Source :
-
DNA and cell biology [DNA Cell Biol] 1990 Dec; Vol. 9 (10), pp. 705-15. - Publication Year :
- 1990
-
Abstract
- Tumor necrosis factor-alpha (TNF-alpha), a protein released by activated macrophages, is involved in a wide variety of human diseases including septic shock, cachexia, and chronic inflammation. TNF binding protein (TNF-BP), a glycoprotein with high affinity to TNF-alpha isolated from urine, acts as an inhibitor of TNF-alpha by competing with the cell-surface TNF receptor. We report here the partial amino acid sequencing of human TNF-BP as well as the isolation, sequence, and expression of cDNA clones encoding a human and rat TNF receptor. The calculated Mr of the mature human and rat TNF receptor chains is 47,526 and 48,072, respectively. The extracellular ligand binding domain represents the soluble TNF-BP which is released by proteolytic cleavage. TNF-BP contains 24 cysteine residues and three potential N-glycosylation sites and shows sequence homology to the extracellular portions of TNF-R p80 chain and nerve growth factor receptor. Transfection of the human TNF receptor cDNA into mammalian cells resulted in increased binding capacity for TNF-alpha and increased reactivity with a monoclonal antibody directed against the human TNF receptor chain p60. When a stop codon was introduced into the cDNA at the site corresponding to the carboxyl terminus of TNF-BP, transfected cells secreted a protein that reacted with antibodies raised against natural TNF-BP.
- Subjects :
- Amino Acid Sequence
Animals
Base Sequence
Blotting, Northern
Cloning, Molecular
DNA genetics
DNA isolation & purification
Humans
Molecular Sequence Data
Plasmids
Poly A analysis
Polymerase Chain Reaction
RNA analysis
RNA, Messenger
Rats
Receptors, Nerve Growth Factor
Receptors, Tumor Necrosis Factor
Transfection
Tumor Necrosis Factor-alpha metabolism
Gene Expression
Receptors, Cell Surface genetics
Tumor Necrosis Factor-alpha genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1044-5498
- Volume :
- 9
- Issue :
- 10
- Database :
- MEDLINE
- Journal :
- DNA and cell biology
- Publication Type :
- Academic Journal
- Accession number :
- 1702293
- Full Text :
- https://doi.org/10.1089/dna.1990.9.705