Secretory synthesis of human interleukin-2 by Streptomyces lividans

Gene. 1990 Feb 14;86(2):227-32. doi: 10.1016/0378-1119(90)90283-w.

Abstract

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.

MeSH terms

  • Base Sequence
  • Cloning, Molecular / methods
  • Humans
  • Immunoblotting
  • Interleukin-2 / biosynthesis
  • Interleukin-2 / genetics*
  • Kinetics
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • Peptides / genetics
  • Plasmids
  • Protein Sorting Signals / genetics
  • Recombinant Fusion Proteins / biosynthesis
  • Restriction Mapping
  • Streptomyces / genetics*
  • alpha-Amylases / antagonists & inhibitors

Substances

  • Interleukin-2
  • Peptides
  • Protein Sorting Signals
  • Recombinant Fusion Proteins
  • alpha-Amylases
  • tendamistate