Rhodostomin, an RGD-containing peptide expressed from a synthetic gene in Escherichia coli, facilitates the attachment of human hepatoma cells.

Abstract

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.

• Authors: H H Chang, S T Hu, T F Huang, S H Chen, Y H Lee, S J Lo
• Journal: Biochemical and biophysical research communications (Biochem Biophys Res Commun) Vol. 190 Issue 1 Pg. 242-9 (Jan 15 1993) ISSN: 0006-291X [Print] United States
• PMID: 7916592 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Oligodeoxyribonucleotides
• Oligopeptides
• Peptides
• Platelet Aggregation Inhibitors
• Recombinant Fusion Proteins
• rhodostomin
• arginyl-glycyl-aspartic acid
• Glutathione Transferase

Topics

• Amino Acid Sequence
• Base Sequence
• Carcinoma, Hepatocellular
• Cell Adhesion (drug effects)
• Cloning, Molecular
• Dose-Response Relationship, Drug
• Escherichia coli (genetics)
• Genes, Synthetic
• Glutathione Transferase (biosynthesis, genetics)
• Humans
• Liver Neoplasms
• Molecular Sequence Data
• Oligodeoxyribonucleotides (chemical synthesis)
• Oligopeptides
• Peptide Biosynthesis
• Peptides (genetics, pharmacology)
• Plasmids
• Platelet Aggregation Inhibitors (pharmacology)
• Recombinant Fusion Proteins (biosynthesis, pharmacology)
• Tumor Cells, Cultured