Prothrombin kringle-2 domain has a growth inhibitory activity against basic fibroblast growth factor-stimulated capillary endothelial cells.
Recently, O'Reilly et al. (O'Reilly, M. S., Holmgren, L., Shing, Y., Chen, C., Rosenthal, R. A., Moses, M., Lane, W. S., Cao, Y., Sage, E. H., and Folkman, J. (1994) Cell 79, 315-328; O'Reilly, M. S., Boehm, T., Shing, Y., Fukai, N., Vasios, G., Lane, W. S., Flynn, E., Birkhead, J. R., Olsen, B. R., and Folkman, J. (1997) Cell 88, 277-285) developed a simple in vitro angiogenesis assay system using bovine capillary endothelial cell proliferation and purified potent angiogenic inhibitors, including angiostatin and endostatin. Using a simple in vitro assay for angiogenesis, we purified a protein molecule that showed anti-endothelial cell proliferative activity from the serum of New Zealand White rabbits, which was stimulated by lipopolysaccharide. The purified protein showed only bovine capillary endothelial cell growth inhibition and not any cytotoxicity. This molecule was identified as a prothrombin kringle-2 domain (fragment-2) using Edman degradation and the amino acid sequence deduced from the cloned cDNA. Both the prothrombin kringle-2 domain released from prothrombin by factor Xa cleavage and the angiogenic inhibitor purified from rabbit sera exhibited anti-endothelial cell proliferative activity. The recombinant rabbit prothrombin kringle-2 domain showed potent inhibitory activity with half-maximal concentrations (ED50) of 2 microg/ml media. As in angiostatin, the recombinant rabbit prothrombin kringle-2 domain also inhibited angiogenesis in the chorioallantoic membrane of chick embryos.
Lee
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Department of Biochemistry, College of Science and Bioproducts Research Center, Yonsei University, Seoul 120-749, Korea.
Rhim
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Kim
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002196:Capillaries / Q000166:cytology*; D002417:Cattle; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D002642:Chick Embryo; D018076:DNA, Complementary; D004730:Endothelium, Vascular / Q000166:cytology*; D016222:Fibroblast Growth Factor 2 / Q000037:antagonists & inhibitors* / Q000502:physiology; D006131:Growth Inhibitors / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008969:Molecular Sequence Data; D011506:Proteins; D011817:Rabbits; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D006131:Growth Inhibitors; D008070:Lipopolysaccharides; D011506:Proteins; C115533:angiogenic inhibitor protein, Oryctolagus cuniculus; D016222:Fibroblast Growth Factor 2