Rhodostomin, an RGD-containing peptide expressed from a synthetic gene in Escherichia coli, facilitates the attachment of human hepatoma cells.
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.
Chang
H H
HH
Graduate Institute of Microbiology and Immunology, National Yang-Ming Medical College, Taipei, Taiwan.
Hu
S T
ST
Huang
T F
TF
Chen
S H
SH
Lee
Y H
YH
Lo
S J
SJ
D000595:Amino Acid Sequence; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular; D002448:Cell Adhesion / Q000187:drug effects*; D003001:Cloning, Molecular; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000235:genetics*; D005813:Genes, Synthetic; D005982:Glutathione Transferase / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D008113:Liver Neoplasms; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis; D009842:Oligopeptides; D010452:Peptide Biosynthesis; D010455:Peptides / Q000235:genetics* / Q000494:pharmacology*; D010957:Plasmids; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D009838:Oligodeoxyribonucleotides; D009842:Oligopeptides; D010455:Peptides; D010975:Platelet Aggregation Inhibitors; D011993:Recombinant Fusion Proteins; C072026:rhodostomin; C047981:arginyl-glycyl-aspartic acid; D005982:Glutathione Transferase