Cloning, expression, and in vitro activity of human endostatin.
Endostatin, a 20 kDa C-terminal fragment of collagen XVIII, is a specific inhibitor of endothelial cell proliferation and angiogenesis. In the present study, we have expressed human endostatin in a yeast expression system (10 mg/L). The recombinant protein was expressed in a soluble form and purified to homogeneity. It specifically inhibited the proliferation and migration of endothelial cells. In addition, we report for the first time that endostatin caused G1 arrest of endothelial cells. Also, we show that endostatin treatment resulted in apoptosis of HUVE and HMVE cells and that all of these effects do not occur in nonendothelial cells. Collectively, these findings demonstrate the expression of a biologically active form of human endostatin in yeast and provide important mechanistic insight into endostatin action on endothelial cells.
D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D001483:Base Sequence; D015153:Blotting, Western; D002417:Cattle; D002453:Cell Cycle; D002455:Cell Division; D002460:Cell Line; D002465:Cell Movement; D003001:Cloning, Molecular; D003094:Collagen / Q000235:genetics* / Q000378:metabolism; D043170:Collagen Type XVIII; D017931:DNA Primers; D043169:Endostatins; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism; D006801:Humans; D010446:Peptide Fragments / Q000235:genetics* / Q000378:metabolism; D010843:Pichia / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism