Protein Engineering vol. 4 no. 3 pp. 335-341, 1991
© 1991 Oxford University Press
RESEARCH-ARTICLE |
The carboxyl-terminal region of human interferon 
is important for biological activity: mutagenic and NMR analysis
Departments of Molecular Biology 60 Orange Street, Bloomfield. NJ 07003, USA 1Departments of Physical Analytical Chemistry Research and Development 60 Orange Street, Bloomfield. NJ 07003, USA 2Departments of Cell Biology, Schering-Plough Research 60 Orange Street, Bloomfield. NJ 07003, USA
Deletion of nine amino acids from the carboxyl terminus of human IFN
(residues 138–146; LFRGRRASQ) resulted in a 7-fold increase in specific antiviral activity. Similar increases in receptor binding affinity were seen. Deletion of residues 136 and 137 (QM) had little additional effect, but removal of Ser135 resulted in a sharp drop in antiviral activity. Further removal of residues 133 and 134 (KR) lowered antiviral activity to 1% of the peak value. Comparison of the proton NMR spectra of selected deletions down to residue 132 showed that there was no significant change in the core protein structure. Deletions down to residue 125 had the same antiviral activity as those to 132, but changes could now be seen in the aromatic proton NMR spectrum of this shorter derivative. Substitution of the homologous murine sequence between residues 124 and 130 (human SPAAKTG; murine LPESSLR) resulted in only a small decrease in antiviral activity, further suggesting that the precise sequence in this region was not critical for activity. Ser135 was substituted with a number of other amino acids with little or no change in activity. The importance of the residues between 131 and 134 for biological activity was corroborated by mutagenesis, although some substitutions in this region were tolerated.
Keywords: interferon /NMR spectroscopy/site-directed mutagenesis/truncation mutants
Received March 12, 1990; revised September 22, 1990; accepted September 22, 1990.
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