Effect of amino-terminal processing by Staphylococcus aureus V-8 protease on activity and structure of recombinant human interferon-gamma.
Treatment of recombinant human interferon-gamma (rHuIFN-gamma) with Staphylococcus aureus V-8 protease generated a transiently stable species that lacks 10 amino-terminal residues. This protein showed distinct secondary and higher-order structures with an alpha-helical content of 31%, suggesting that the secondary and tertiary structure largely remain upon removal of 10 amino-terminal residues. The antiviral activity was abolished, or greatly reduced, for this species relative to the intact protein. These results suggest an important role for the amino-terminal portion in the activity of human interferon-gamma. Since the digested protein is difficult to refold from the acid-denatured state, it was concluded that, although not essential for maintaining the core structure of the protein, the amino-terminal portion is critical for refolding the protein from acid.
Arakawa
T
T
Amgen Inc., Thousand Oaks, CA 91320.
Horan
T P
TP
McGinley
M
M
Rohde
M F
MF
D000595:Amino Acid Sequence; D006801:Humans; D007371:Interferon-gamma / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000302:isolation & purification; D011487:Protein Conformation; D011994:Recombinant Proteins; D012697:Serine Endopeptidases / Q000494:pharmacology*; D013211:Staphylococcus aureus / Q000201:enzymology; D013329:Structure-Activity Relationship
D010446:Peptide Fragments; D011994:Recombinant Proteins; D007371:Interferon-gamma; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase