Identification of the binding site of 55kda tumor necrosis factor receptor by synthetic peptides

• Published in: Biochemical and biophysical research communications Vol. 188; no. 2; pp. 503 - 509
• Main Authors: Lie, Byong Lyul, Tunemoto, Daiei, Hemmi, Hiromichi, Mizukami, Yukiko, Fukuda, Hiroko, Kikuchi, Hiroshi, Kato, Seishi, Numao, Naganori
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 30.10.1992; Elsevier
• Subjects: Amino Acid Sequence; Binding Sites; Binding, Competitive; Biological and medical sciences; Cell Line; Cell receptors; Cell structures and functions; Cytokines - pharmacology; Fundamental and applied biological sciences. Psychology; Humans; Interferon-gamma - pharmacology; Interleukin-1 - pharmacology; Interleukin-6 - pharmacology; Kinetics; Miscellaneous; Molecular and cellular biology; Molecular Sequence Data; Molecular Weight; Peptides - chemical synthesis; Peptides - metabolism; Receptors, Cell Surface - chemistry; Receptors, Cell Surface - drug effects; Receptors, Cell Surface - metabolism; Receptors, Tumor Necrosis Factor; Recombinant Proteins - metabolism; Recombinant Proteins - pharmacology; Tumor Necrosis Factor-alpha - metabolism; Tumor Necrosis Factor-alpha - pharmacology; Human; Membrane receptor; Binding site; Tumor necrosis factor; Cytokine
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/0006-291X(92)91084-4

We have synthesized a series of peptides, which cover almost the whole range of the N-terminal extracellular domain of human 55kDa TNF receptor (55kDa TNF-R). The peptides were examined for the binding activity to TNF by solid phase binding assay and for the inhibition of TNF cytotoxicity to mouse L-M cells. The peptide 159–178 exhibited remarkably higher binding activity to TNF than other peptides did. The specificity of the TNF binding to the peptides was confirmed by their inability to bind other cytokines. The peptide 159–178 also inhibited TNF cytotoxicity. These results indicate that the specific binding site of 55kDa TNF-R to TNF might reside within the peptide segment of amino acid numbers 159 to 178 in the N-terminal extracellular domain.