Synthesis, antiviral activity, and conformational characterization of mouse-human alpha-interferon hybrids.
Reciprocal hybrids were constructed between human and mouse interferons (IFNs), and their antiviral activity was examined on different target cells and compared to the activity of the parental molecules. In addition, we used a number of predictive algorithms on a data base of the available alpha-interferon sequences to propose a working model for the overall conformation of the alpha-interferon molecule that is consistent with the structural predictions. Remarkable conservation within the predicted alpha-helical segments of the interferon molecule was observed. We propose that the observed changes in the activity and specificity of the hybrids obtained are largely due to the sequences present in the loops at the ends of the major helical structures; these are less conserved, contain beta-bends, and are generally hydrophilic and flexible. The data on the constructed mouse-human hybrids have shown that the activity on human cells is contributed by determinants present in the N-terminal 122 amino acids of human IFN, thus implicating one or more loops within this region (e.g. loops 1-12, 25-38, 70-74, and 103-113). The activity on bovine cells appears to be localized mainly in sequence 60-121, implicating the role of loops 70-74 and/or 103-113 of the human IFN molecule. The specificity of mouse IFN for mouse cells is in some or all of the loops (70-74, 103-113, 134-139, and 163-166) in the C-terminal sequence. The proposed working model should provide guidelines for the study of the specificity of action in molecular terms.
Raj
N B
NB
Oncology Center, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Israeli
R
R
Kelley
K A
KA
Leach
S J
SJ
Minasian
E
E
Sikaris
K
K
Parry
D A
DA
Pitha
P M
PM
D000595:Amino Acid Sequence; D000818:Animals; D004247:DNA / Q000235:genetics; D004262:DNA Restriction Enzymes; D004686:Encephalomyocarditis virus / Q000187:drug effects*; D004926:Escherichia coli / Q000235:genetics; D005796:Genes; D006801:Humans; D007370:Interferon Type I / Q000235:genetics* / Q000494:pharmacology; D051379:Mice; D008826:Microbial Sensitivity Tests; D008958:Models, Molecular; D010957:Plasmids; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000494:pharmacology; D013045:Species Specificity; D014721:Vesicular stomatitis Indiana virus / Q000187:drug effects*
D007370:Interferon Type I; D011994:Recombinant Proteins; D004247:DNA; D004262:DNA Restriction Enzymes