High level synthesis of biologically active recombinant trichosanthin in Escherichia coli

Int J Pept Protein Res. 1992 Jan;39(1):77-81. doi: 10.1111/j.1399-3011.1992.tb01558.x.

Abstract

Two forms of recombinant trichosanthin (rTCS) were synthesized in high levels in Escherichia coli by putting the TCS cDNA under the control of a T7 RNA polymerase-directed promoter. Purification schemes were developed to isolate the recombinant protein from both soluble and insoluble fractions. Form I rTCS possessed the mature TCS sequence and had similar biological activities as the natural protein. Its IC50 was approximately 0.13 nM in an in vitro rabbit reticulocyte translational system and a dose of around 35 micrograms protein per 25 g body weight was sufficient to induce complete abortion in mice. Form II rTCS had a propeptide of 19 aa at the C-terminus and was five times less active than Form I in inhibiting protein synthesis by a rabbit reticulocyte lysate.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Abortifacient Agents / pharmacology
  • Amino Acid Sequence
  • Animals
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Female
  • Molecular Sequence Data
  • Protein Biosynthesis / drug effects
  • Rabbits
  • Recombinant Proteins / biosynthesis*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / pharmacology
  • Ribosomes / drug effects
  • Solubility
  • Trichosanthin / biosynthesis*
  • Trichosanthin / chemistry
  • Trichosanthin / isolation & purification
  • Trichosanthin / pharmacology

Substances

  • Abortifacient Agents
  • Recombinant Proteins
  • Trichosanthin