Production and characterization of a monoclonal antibody to a human interferon-induced double-stranded RNA-binding Mr 68,000 protein kinase.
One of the interferon-induced proteins thought to be involved in the antiviral effects of interferon is a double-stranded RNA-dependent protein kinase. This paper reports the development of a monoclonal antibody, 10A5, that recognizes a protein that co-migrates with the double-stranded RNA-dependent protein kinase at an approximate molecular weight of 68,000. Levels of this protein and of the protein kinase activity increase 3-fold on interferon treatment of T98G cells. The specificity of the monoclonal antibody was determined by ELISA, immunoblotting, and immunoprecipitation procedures. Furthermore, immunoaffinity chromatography of an interferon-induced T98G cell extract previously phosphorylated in the presence of double-stranded RNA and radiolabeled ATP resulted in the specific elution of a phosphorylated Mr 68,000 protein from the monoclonal antibody 10A5-Sepharose column. Monoclonal antibody 10A5 recognizes both native and denatured protein kinase, independent of double-stranded RNA binding or phosphorylation, and should therefore serve as a useful tool in analyzing the role of the double-stranded RNA-dependent protein kinase in the mechanism of interferon action.
D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000918:Antibody Specificity; D002846:Chromatography, Affinity; D004790:Enzyme Induction / Q000187:drug effects; D006801:Humans; D007370:Interferon Type I / Q000494:pharmacology*; D008970:Molecular Weight; D011070:Poly I-C / Q000378:metabolism; D011494:Protein Kinases / Q000276:immunology*; D012330:RNA, Double-Stranded / Q000378:metabolism*; D019892:eIF-2 Kinase
D000911:Antibodies, Monoclonal; D007370:Interferon Type I; D012330:RNA, Double-Stranded; D011494:Protein Kinases; D019892:eIF-2 Kinase; D011070:Poly I-C