Specific binding of human alpha interferon to a high affinity cell surface binding site on bovine kidney cells.
Virus-induced human alpha interferon (HuIFN-alpha) derived from Namalwa cells and purified to a specific activity of 2 X 10(8) units/mg of protein was radiolabeled with 125I-labeled Bolton and Hunter reagent to a specific activity of 4-12 microCi/micrograms of protein. The binding of this 125I-IFN to bovine kidney cells was examined at 4 degrees C. Scatchard analysis of the binding data indicate the presence of 650 binding sites/cell and binding of the ligand with an apparent Kd of 6 X 10(-11) M. Trypsin or acid treatment of cells to which 125I-IFN was bound resulted in the release of greater than or equal to 77% of the radioactivity, indicating a majority of radiolabeled material was bound to the cell surface. Antibodies against human leukocyte IFN but not antibodies against human fibroblast IFN inhibited the binding of radiolabeled IFN to the cells. The binding of 125I-IFN was not inhibited by a 75-fold molar excess of mouse IFN but was inhibited 30% by a 200-fold molar excess of human beta (fibroblast) IFN. These data are compatible with the Lower biological activities of these IFNs on bovine kidney cells. Several Escherichia coli derived HuIFN-alpha s inhibited the binding of the radiolabeled IFN to the same extent as native HuIFN-alpha s, but four fragments of HuIFN-alpha 1, an E. coli-derived 86 amino acid NH2-terminal fragment as well as 3 different synthetic carboxy-terminal fragments of 140, 56, or 46 amino acids did not inhibit binding.
D000818:Animals; D000936:Antigen-Antibody Complex; D001667:Binding, Competitive; D002417:Cattle; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D006801:Humans; D007372:Interferons / Q000276:immunology / Q000378:metabolism*; D007668:Kidney / Q000378:metabolism*; D007700:Kinetics; D011956:Receptors, Cell Surface / Q000378:metabolism*; D017471:Receptors, Interferon; D005719:gamma-Globulins
D000936:Antigen-Antibody Complex; D011956:Receptors, Cell Surface; D017471:Receptors, Interferon; D005719:gamma-Globulins; D007372:Interferons