Production and structural characterization of amino terminally histidine tagged human oncostatin M in E. coli

Cytokine. 1994 May;6(3):255-64. doi: 10.1016/1043-4666(94)90021-3.


Oncostatin M is a cytokine that acts as a growth regulator on a wide variety of cells and has diverse biological activities including acute phase protein induction, LDL receptor up-regulation and cell-specific gene expression. In order to gather information about the Onc M structure, we established a protocol for large scale production and single step purification of this functional cytokine from bacterial cells. The cDNA of human Onc M was cloned by RT-PCR from total RNA of PMA induced U937 cells. After the addition of a six histidine tag at the N-terminus, the coding region of mature Onc M was cloned in the pT7.7 expression vector. Histidine tagged Onc M was overexpressed in bacterial cells and purified to homogeneity in one step on a metal chelating column. We found that recombinant 6xHis-OncM remains fully active in a growth inhibition assay. Structural characterization of the purified protein was performed by electrospray mass spectrometry, automated Edman degradation and peptide mapping by high-pressure liquid chromatography/fast-atom-bombardment mass spectrometry. Thermal and pH stability dependence of Onc M was assessed by circular dichroism spectroscopy; the helical content is about 50%, in agreement with the four helix bundle fold postulated for cytokines that bind haematopoietic receptors of type I.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cell Line
  • Chromatography, High Pressure Liquid
  • Cloning, Molecular / methods
  • Cytokines / biosynthesis
  • Escherichia coli
  • Gene Expression
  • Growth Inhibitors / biosynthesis*
  • Histidine*
  • Humans
  • Models, Structural
  • Molecular Sequence Data
  • Oncostatin M
  • Peptide Biosynthesis*
  • Peptides / chemistry*
  • Polymerase Chain Reaction
  • Protein Structure, Secondary*
  • Recombinant Proteins / biosynthesis*
  • Recombinant Proteins / chemistry
  • Spectrometry, Mass, Fast Atom Bombardment
  • Tetradecanoylphorbol Acetate / pharmacology
  • Transcription, Genetic


  • Cytokines
  • Growth Inhibitors
  • OSM protein, human
  • Peptides
  • Recombinant Proteins
  • Oncostatin M
  • Histidine
  • Tetradecanoylphorbol Acetate