rhodostomin and Carcinoma--Hepatocellular

Rhodostomin (Rho) from snake venom, a potent inhibitor of platelet aggregation, contains 68 amino acids having an RGD sequence and 12 cysteine residues. A chemically synthesized Rho gene was cloned and expressed in Escherichia coli. The expression of Rho gene fused with the glutathione S-transferase (GST) gene was about 10-30% of total cell proteins. The Rho-fusion protein could be recognized by antibodies raised against either a native Rho peptide or a synthetic peptide. The purified GST-Rho coated on culture plates facilitated the attachment of human hepatoma cells, which was inhibitable by co-incubation with a synthetic hexapeptide GRGDSP but not with a related peptide of GRGESP, suggesting that the E. coli-expressed Rho-fusion protein was properly folded and biologically functional.

Topics: Amino Acid Sequence; Base Sequence; Carcinoma, Hepatocellular; Cell Adhesion; Cloning, Molecular; Dose-Response Relationship, Drug; Escherichia coli; Genes, Synthetic; Glutathione Transferase; Humans; Liver Neoplasms; Molecular Sequence Data; Oligodeoxyribonucleotides; Oligopeptides; Peptide Biosynthesis; Peptides; Plasmids; Platelet Aggregation Inhibitors; Recombinant Fusion Proteins; Tumor Cells, Cultured