This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Olsson, A.-K. Articles by Claesson-Welsh, L. Search for Related Content PubMed PubMed Citation Articles by Olsson, A.-K. Articles by Claesson-Welsh, L.

The Minimal Active Domain of Endostatin Is a Heparin-Binding Motif that Mediates Inhibition of Tumor Vascularization

¹ Department of Genetics and Pathology, Uppsala University, Rudbeck Laboratory, Uppsala, Sweden; ² Department of Medical Cell Biology, Biomedical Centre, Uppsala, Sweden; and ³ Laboratory of Protein Chemistry, Max-Planck Institute for Biochemistry, Martinsried, Germany

Endostatin constitutes the COOH-terminal 20,000 Da proteolytic fragment of collagen XVIII and has been shown to possess antiangiogenic and antitumorigenic properties. In the present study, we have investigated the role of the heparin-binding sites in the in vivo mechanism of action of endostatin. The majority of the heparin binding is mediated by arginines 155/158/184/270 in endostatin, but there is also a minor site constituted by arginines 193/194. Using endostatin mutants lacking either of these two sites, we show that inhibition of fibroblast growth factor-2–induced angiogenesis in the chicken chorioallantoic membrane requires both heparin-binding sites. In contrast, inhibition of vascular endothelial growth factor-A–induced chorioallantoic membrane angiogenesis by endostatin was only dependent on the minor heparin-binding site (R193/194). These arginines were also required for endostatin to inhibit fibroblast growth factor-2– and vascular endothelial growth factor-A–induced chemotaxis of primary endothelial cells. Moreover, we show that a synthetic peptide corresponding to amino acids 180–199 of human endostatin (which covers the minor heparin-binding site) inhibits endothelial cell chemotaxis and reduces tumor vascularization in vivo. Substitution of arginine residues 193/194 for alanine attenuates the antiangiogenic effects of the peptide. These data show an essential role for heparin binding in the antiangiogenic action of endostatin.

This article has been cited by other articles:

				M. H. Kulke, E. K. Bergsland, D. P. Ryan, P. C. Enzinger, T. J. Lynch, A. X. Zhu, J. A. Meyerhardt, J. V. Heymach, W. E. Fogler, C. Sidor, et al. Phase II Study of Recombinant Human Endostatin in Patients With Advanced Neuroendocrine Tumors J. Clin. Oncol., August 1, 2006; 24(22): 3555 - 3561. [Abstract] [Full Text] [PDF]

				M. Vanwildemeersch, A.-K. Olsson, E. Gottfridsson, L. Claesson-Welsh, U. Lindahl, and D. Spillmann The Anti-angiogenic His/Pro-rich Fragment of Histidine-rich Glycoprotein Binds to Endothelial Cell Heparan Sulfate in a Zn2+-dependent Manner J. Biol. Chem., April 14, 2006; 281(15): 10298 - 10304. [Abstract] [Full Text] [PDF]

				J. Dixelius, A.-K. Olsson, A. Thulin, C. Lee, I. Johansson, and L. Claesson-Welsh Minimal Active Domain and Mechanism of Action of the Angiogenesis Inhibitor Histidine-Rich Glycoprotein Cancer Res., February 15, 2006; 66(4): 2089 - 2097. [Abstract] [Full Text] [PDF]

				S. Stahl, S. Gaetzner, T. D. Mueller, and U. Felbor Endostatin phenylalanines 31 and 34 define a receptor binding site Genes Cells, September 1, 2005; 10(9): 929 - 939. [Abstract] [Full Text] [PDF]