Expression and secretion of biologically active echistatin in Saccharomyces cerevisiae

Gene. 1989 Dec 28;85(2):511-6. doi: 10.1016/0378-1119(89)90445-9.

Abstract

A synthetic gene coding for a platelet aggregation inhibitor, echistatin (ECS), was inserted into a Saccharomyces cerevisiae expression vector utilizing the alpha-mating factor pre-pro leader sequence and galactose-inducible promoter, GAL10. Cleavage of the pre-pro leader sequence in vivo results in the secretion of a properly processed recombinant ECS with the native N-terminal glutamic acid residue. Recombinant ECS was recovered from yeast supernatants and purified by reverse phase high performance liquid chromatography. Recombinant ECS expressed and purified from yeast was identical to native ECS in its ability to inhibit platelet aggregation.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Endopeptidases / genetics
  • Genes, Synthetic*
  • Genetic Vectors
  • Humans
  • Intercellular Signaling Peptides and Proteins
  • Mating Factor
  • Membrane Proteins*
  • Molecular Sequence Data
  • Peptides / genetics
  • Pheromones / biosynthesis
  • Platelet Aggregation
  • Platelet Aggregation Inhibitors / metabolism*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / pharmacology
  • Restriction Mapping
  • Saccharomyces cerevisiae / genetics*
  • Serine Endopeptidases*
  • Viper Venoms / biosynthesis
  • Viper Venoms / genetics*
  • Viper Venoms / pharmacology

Substances

  • Intercellular Signaling Peptides and Proteins
  • Membrane Proteins
  • Peptides
  • Pheromones
  • Platelet Aggregation Inhibitors
  • Recombinant Proteins
  • Viper Venoms
  • echistatin
  • Mating Factor
  • Endopeptidases
  • Serine Endopeptidases
  • type I signal peptidase