The Tumor-Suppressing Activity of Angiostatin Protein Resides within Kringles 1 to 3

Abstract

Angiostatin protein, which comprises the first four kringle domains of plasminogen, is an endogenous inhibitor of angiogenesis that inhibits the growth of experimental primary and metastatic tumors. Truncation of Angiostatin K1–4 to K1–3 retained the activity of Angiostatin. We recombinantly expressed full-length human Angiostatin protein corresponding to the first four kringle domains of human plasminogen and a truncated form of the Angiostatin protein, kringles 1–3. Purified recombinant Angiostatin K1–3 and K1–4 proteins inhibited the formation of experimental B16-BL6 lung metastases by greater than 80% when administered at 30 nmol/kg/day. We demonstrate for the first time that Angiostatin protein, consisting of the first three kringle domains of human plasminogen, has in vivo biological activity in this assay indistinguishable from that of the full-length Angiostatin K1–4 protein and that the fourth kringle of plasminogen, when linked in sequence to K1–3, plays no direct role in the antitumor activity of Angiostatin.