Structure/function studies on recombinant human gamma interferon.
Structure/function relationships for human gamma interferon (IFN-gamma) were investigated using recombinant DNA-derived homologues produced in E. coli. The various biological effects examined were antiviral, growth inhibitory and 2-5A synthetase activities, as well as receptor binding characteristics. Specific structural changes led to IFN-gamma homologues with defined alterations in biological activities. Amino acid residue changes at the hydrophobic core of the molecule resulted in two homologues exhibiting loss of affinity for the IFN-gamma receptor and dramatically reduced biological activities. These diminished activities probably relate to the inability of the homologues to form appropriately folded structures. Residue changes at two sites associated with beta-turns on the surface of IFN-gamma likewise resulted in homologues with reduced biological activities. In these cases, the reduced biological activities were not associated with reduced receptor binding. Addition of cysteine-tyrosine-cysteine to the amino-terminus of IFN-gamma, known to perturb the protein conformation, slightly reduced the affinity of the so-derived homologue for the IFN-gamma receptor on T98G cells, and there was concommitant reduction in biological activities. Experiments with a monoclonal antibody that binds to the carboxy-terminus of IFN-gamma indicated that this region of the molecule may not influence antiviral or antiproliferative activities. Overall our data imply that several sites along the IFN-gamma polypeptide contribute to biological activity, and that receptor binding and effector sites are distinct.
Fish
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EN
Research Institute, Hospital for Sick Children, Toronto, Canada.
Banerjee
K
K
Arakawa
T
T
Stebbing
N
N
D015088:2',5'-Oligoadenylate Synthetase / Q000096:biosynthesis*; D000595:Amino Acid Sequence; D000998:Antiviral Agents / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects*; D002460:Cell Line; D004686:Encephalomyocarditis virus / Q000187:drug effects; D004790:Enzyme Induction; D004926:Escherichia coli / Q000235:genetics; D005796:Genes; D006801:Humans; D007371:Interferon-gamma / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D007700:Kinetics; D008214:Lymphocytes / Q000276:immunology; D009154:Mutation; D011971:Receptors, Immunologic / Q000378:metabolism*; D017471:Receptors, Interferon; D011994:Recombinant Proteins; D013329:Structure-Activity Relationship
D000998:Antiviral Agents; D011971:Receptors, Immunologic; D017471:Receptors, Interferon; D011994:Recombinant Proteins; D007371:Interferon-gamma; D015088:2',5'-Oligoadenylate Synthetase