Secretory synthesis of human interleukin-2 by Streptomyces lividans.
To study the ability of Streptomyces lividans to produce heterologous proteins by secretion, we directly fused DNA encoding the leader peptide of the alpha-amylase inhibitor, tendamistat, produced by Streptomyces tendae, with DNA encoding the mature part of interleukin-2 (IL-2). Such cloned fusion constructs are translated in S. lividans, in spite of the quite different codon usage. The active Il-2 is secreted into the culture broth, though the amounts are much less than that of the alpha-amylase inhibitor. The presence of IL-2 in the supernatants could be demonstrated both by an activity assay and by immunoblotting. In addition to the secreted form, three different species of Il-2 antibody immunoreactive proteins, with different Mrs, are either present in the cells or attached to the cells. This indicates that inefficient processing and translocation of the precursor is a major reason for the low activities found in the supernatant.
Bender
E
E
Institut für Organische Chemie, Universität Frankfurt, F.R.G.
Koller
K P
KP
Engels
J W
JW
D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D006801:Humans; D015151:Immunoblotting; D007376:Interleukin-2 / Q000096:biosynthesis / Q000235:genetics*; D007700:Kinetics; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010455:Peptides / Q000235:genetics; D010957:Plasmids; D021382:Protein Sorting Signals / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D015183:Restriction Mapping; D013302:Streptomyces / Q000235:genetics*; D000516:alpha-Amylases / Q000037:antagonists & inhibitors
D007376:Interleukin-2; D010455:Peptides; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D000516:alpha-Amylases; C030526:tendamistate