The human interferon-gamma receptor. Purification, characterization, and preparation of antibodies.
The receptor for human interferon-gamma (IFN-gamma) was purified from foreskin fibroblasts. Triton X-100 extracts obtained from either intact cells or membrane preparations were passed through an immobilized interferon-gamma column. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of eluted fractions revealed a major band of Mr = 95,000 and minor bands of Mr = 80,000 and 60,000. Further purification was obtained by steric exclusion and by lectin chromatography. The purified receptor retained the ability to bind 125I-IFN-gamma with a Kd of 2.2 X 10(-10) M, a value close to that obtained with intact fibroblasts (5 X 10(-10) M). A complex of Mr = 105,000-125,000 was visualized by immunoprecipitation of 125I-IFN-gamma cross-linked to the purified receptor followed by SDS-PAGE and autoradiography. A similar complex was obtained when 125I-IFN-gamma was cross-linked to intact cells. Immunization of mice with the excised SDS-PAGE band of Mr = 95,000 elicited antibodies that blocked the antiviral activity of IFN-gamma and immunoprecipitated the cross-linked complex of 125I-IFN-gamma and its receptor.
D000906:Antibodies; D002462:Cell Membrane / Q000276:immunology; D002846:Chromatography, Affinity / Q000379:methods; D005347:Fibroblasts / Q000276:immunology; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D008970:Molecular Weight; D011971:Receptors, Immunologic / Q000302:isolation & purification* / Q000378:metabolism; D017471:Receptors, Interferon; D012867:Skin / Q000276:immunology
D000906:Antibodies; D011971:Receptors, Immunologic; D017471:Receptors, Interferon; D007371:Interferon-gamma