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Angiopoietins have distinct modular domains essential for receptor binding, dimerization and superclustering

Nature Structural Biology volume 10, pages 38–44 (2003)Cite this article

A Corrigendum to this article was published on 01 February 2003

Abstract

Angiopoietins are a recently discovered family of angiogenic factors that interact with the endothelial receptor tyrosine kinase Tie2, either as agonists (angiopoietin-1) or as context-dependent agonists/antagonists (angiopoietin-2). Here we show that angiopoietin-1 has a modular structure unlike any previously characterized growth factor. This modular structure consists of a receptor-binding domain, a dimerization motif and a superclustering motif that forms variable-sized multimers. Genetic engineering of precise multimers of the receptor-binding domain of angiopoietin-1, using surrogate multimerization motifs, reveals that tetramers are the minimal size required for activating endothelial Tie2 receptors. In contrast, engineered dimers can antagonize endothelial Tie2 receptors. Surprisingly, angiopoietin-2 has a modular structure and multimerization state similar to that of angiopoietin-1, and its antagonist activity seems to be a subtle property encoded in its receptor-binding domain.

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Figure 1: Structure and activity of Ang1 and Ang1 variants.
Figure 2: Analysis of multimericity, receptor-binding affinity and receptor-binding stoichiometry of angiopoietin variants.
Figure 3: Effects of angiopoietin multimericity and F domain chimeras on Tie2 activation.
Figure 4: Transmission electron microscopy visualization of angiopoietin oligomers.
Figure 5: Engineered tetramers of Ang1 are useful in vivo reagents.

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References

  1. Davis, S. et al. Isolation of angiopoietin-1, a ligand for the TIE2 receptor, by secretion-trap expression cloning. Cell 87, 1161–1169 (1996).

    Article  CAS  Google Scholar 

  2. Suri, C. et al. Requisite role of angiopoietin-1, a ligand for the TIE2 receptor, during embryonic angiogenesis. Cell 87, 1171–1180 (1996).

    Article  CAS  Google Scholar 

  3. Maisonpierre, P.C. et al. Angiopoietin-2, a natural antagonist for Tie2 that disrupts in vivo angiogenesis. Science 277, 55–60 (1997).

    Article  CAS  Google Scholar 

  4. Valenzuela, D.M. et al. Angiopoietins 3 and 4: diverging gene counterparts in mice and humans. Proc. Natl. Acad. Sci. USA 96, 1904–1909 (1999).

    Article  CAS  Google Scholar 

  5. Yancopoulos, G.D. et al. Vascular-specific growth factors and blood vessel formation. Nature 407, 242–248 (2000).

    Article  CAS  Google Scholar 

  6. Gale, N.W. et al. Angiopoietin-2 is required for postnatal angiogenesis and lymphatic patterning, and only the latter role is rescued by angiopoietin-1. Dev. Cell 3, 411–423 (2002).

    Article  CAS  Google Scholar 

  7. Jones, N., Iljin, K., Dumont, D.J. & Alitalo, K. Tie receptors: new modulators of angiogenic and lymphangiogenic responses. Nat. Rev. Mol. Cell. Biol. 2, 257–267 (2001).

    Article  CAS  Google Scholar 

  8. Teichert-Kuliszewska, K. et al. Biological action of angiopoietin-2 in a fibrin matrix model of angiogenesis is associated with activation of Tie2. Cardiovasc. Res. 49, 659–670 (2001).

    Article  CAS  Google Scholar 

  9. Mochizuki, Y., Nakamura, T., Kanetake, H. & Kanda, S. Angiopoietin 2 stimulates migration and tube-like structure formation of murine brain capillary endothelial cells through c-Fes and c-Fyn. J. Cell Sci. 115, 175–183 (2002).

    CAS  PubMed  Google Scholar 

  10. Cunningham, B.C. et al. Dimerization of the extracellular domain of the human growth hormone receptor by a single hormone molecule. Science 254, 821–825 (1991).

    Article  CAS  Google Scholar 

  11. de Vos, A.M., Ultsch, M. & Kossiakoff, A.A. Human growth hormone and extracellular domain of its receptor: crystal structure of the complex. Science 255, 306–312 (1992).

    Article  CAS  Google Scholar 

  12. Schlessinger, J., Lax, I. & Lemmon, M. Regulation of growth factor activation by proteoglycans: what is the role of the low affinity receptors? Cell 83, 357–360 (1995).

    Article  CAS  Google Scholar 

  13. Davis, S. et al. Ligands for EPH-related receptor tyrosine kinases that require membrane attachment or clustering for activity. Science 266, 816–819 (1994).

    Article  CAS  Google Scholar 

  14. Procopio, W.N., Pelavin, P.I., Lee, W.M. & Yeilding, N.M. Angiopoietin-1 and -2 coiled coil domains mediate distinct homo-oligomerization patterns, but fibrinogen-like domains mediate ligand activity. J. Biol. Chem. 274, 30196–30201 (1999).

    Article  CAS  Google Scholar 

  15. Doolittle, R.F. Fibrinogen and fibrin. Annu. Rev. Biochem. 53, 195–229 (1984).

    Article  CAS  Google Scholar 

  16. Wen, J., Arakawa, T. & Philo, J.S. Size-exclusion chromatography with on-line light-scattering, absorbance, and refractive index detectors for studying proteins and their interactions. Anal. Biochem. 240, 155–166 (1996).

    Article  CAS  Google Scholar 

  17. Suri, C. et al. Increased vascularization in mice overexpressing angiopoietin-1. Science 282, 468–471 (1998).

    Article  CAS  Google Scholar 

  18. Thurston, G. et al. Leakage-resistant blood vessels in mice transgenically overexpressing angiopoietin-1. Science 286, 2511–2514 (1999).

    Article  CAS  Google Scholar 

  19. Thurston, G. et al. Angiopoietin-1 protects the adult vasculature against plasma leakage. Nat. Med. 6, 460–463 (2000).

    Article  CAS  Google Scholar 

  20. Karlsson, R., Michaelsson, A. & Mattsson, L. Kinetic analysis of monoclonal antibody-antigen interactions with a new biosensor based analytical system. J. Immunol. Methods 145, 229–240 (1991).

    Article  CAS  Google Scholar 

  21. Johnsson, B., Lofas, S. & Lindquist, G. Immobilization of proteins to a carboxymethyldextran-modified gold surface for biospecific interaction analysis in surface plasmon resonance sensors. Anal. Biochem. 198, 268–277 (1991).

    Article  CAS  Google Scholar 

  22. Edgell, C.J., McDonald, C.C. & Graham, J.B. Permanent cell line expressing human factor VIII-related antigen established by hybridization. Proc. Natl. Acad. Sci. USA 80, 3734–3737 (1983).

    Article  CAS  Google Scholar 

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Acknowledgements

We gratefully acknowledge D. Datta, K. Lamb, A. Polotskaia, and S. Xu for excellent technical assistance, and N. Stahl for insightful conversations and advice.

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  1. Regeneron Pharmaceuticals, Inc., 777 Old Saw Mill River Road, Tarrytown, 10591, New York, USA

    Samuel Davis, Nick Papadopoulos, Thomas H. Aldrich, Peter C. Maisonpierre, Tammy Huang, Lubomir Kovac, April Xu, Raymond Leidich, Elzbieta Radziejewska, Ashique Rafique, Judah Goldberg, Vivek Jain, Kevin Bailey, Margaret Karow, Jim Fandl, Ella Ioffe, John S. Rudge, Thomas J. Daly, Czeslaw Radziejewski & George D. Yancopoulos

  2. Procter and Gamble Pharmaceuticals, P.O. Box 8006, 8700 Mason-Montgomery Road, Mason, 45040-8006, Ohio, USA

    Steven J. Samuelsson

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Correspondence to Samuel Davis.

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The authors are employees of Regeneron Pharmaceuticals, Inc., or Procter and Gamble Pharmaceuticals.

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Davis, S., Papadopoulos, N., Aldrich, T. et al. Angiopoietins have distinct modular domains essential for receptor binding, dimerization and superclustering. Nat Struct Mol Biol 10, 38–44 (2003). https://doi.org/10.1038/nsb880

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