Amino acid sequence analysis, gene construction, cloning, and expression of gelonin, a toxin derived from Gelonium multiflorum

J Interferon Cytokine Res. 1995 Jun;15(6):547-55. doi: 10.1089/jir.1995.15.547.

Abstract

The plant toxin gelonin is an extremely potent inhibitor of protein synthesis, similar in action to ricin. The mature protein primary sequence was obtained using conventional sequencing techniques. Gelonin was found to be composed of 258 amino acids and contains 21 lysine residues. This toxin shares approximately 33% sequence homology with trichosanthin and ricin A chain. A 774 bp synthetic gene encoding gelonin was synthesized and expressed in E. coli. Recombinant gelonin (approximately 28 kD) expression was monitored and demonstrated by western analysis. Purification and functional activity studies demonstrated that this protein behaves identically to that of the natural product. Recombinant gelonin (RG) thus joins a growing list of recombinant toxins currently available for use in the construction of recombinant immunotoxins composed of gelonin fused to binding domains of antibodies, growth factors, or other cytokines.

MeSH terms

  • Amino Acid Sequence
  • Cloning, Molecular
  • Gene Expression
  • Molecular Sequence Data
  • Plant Proteins / biosynthesis
  • Plant Proteins / genetics*
  • Plant Proteins / isolation & purification
  • Protein Synthesis Inhibitors / isolation & purification
  • Protein Synthesis Inhibitors / metabolism*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / isolation & purification
  • Ribosome Inactivating Proteins, Type 1
  • Sequence Homology, Amino Acid
  • Toxins, Biological / biosynthesis
  • Toxins, Biological / genetics*
  • Toxins, Biological / isolation & purification

Substances

  • Plant Proteins
  • Protein Synthesis Inhibitors
  • Recombinant Proteins
  • Ribosome Inactivating Proteins, Type 1
  • Toxins, Biological
  • GEL protein, Gelonium multiflorum