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Crystal structures of the Tie2 receptor ectodomain and the angiopoietin-2–Tie2 complex

Nature Structural & Molecular Biology volume 13, pages 524–532 (2006)Cite this article

Abstract

The Tie receptor tyrosine kinases and their angiopoietin (Ang) ligands play central roles in developmental and tumor-induced angiogenesis. Here we present the crystal structures of the Tie2 ligand-binding region alone and in complex with Ang2. In contrast to prediction, Tie2 contains not two but three immunoglobulin (Ig) domains, which fold together with the three epidermal growth factor domains into a compact, arrowhead-shaped structure. Ang2 binds at the tip of the arrowhead utilizing a lock-and-key mode of ligand recognition—unique for a receptor kinase—where two complementary surfaces interact with each other with no domain rearrangements and little conformational change in either molecule. Ang2-Tie2 recognition is similar to antibody–protein antigen recognition, including the location of the ligand-binding site within the Ig fold. Analysis of the structures and structure-based mutagenesis provide insight into the mechanism of receptor activation and support the hypothesis that all angiopoietins interact with Tie2 in a structurally similar manner.

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Figure 1: Structure of the Tie2 ligand-binding region.
Figure 2: Sequence alignment of the interacting domains of angiopoietins and Ties.
Figure 3: Ligand binding and activation of wild-type and mutant Tie2.
Figure 4: Crystal structure of the Ang2–Tie2 complex.
Figure 5: Structure of the Ang2-Tie2 interface.
Figure 6: A model for Tie2 activation by angiopoietins.

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References

  1. Folkman, J. & D'Amore, P.A. Blood vessel formation: What is its molecular basis? Cell 87, 1153–1155 (1996).

    Article  CAS  PubMed  Google Scholar 

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

    CAS  PubMed  Google Scholar 

  3. 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  PubMed  Google Scholar 

  4. Carmeliet, P. & Jain, R.K. Angiogenesis in cancer and other diseases. Nature 407, 249–257 (2000).

    Article  CAS  PubMed  Google Scholar 

  5. Sato, T.N. Vascular development: molecular logic for defining arteries and veins. Curr. Opin. Hematol. 10, 131–135 (2003).

    Article  CAS  PubMed  Google Scholar 

  6. Semenza, G.L. Angiogenesis in ischemic and neoplastic disorders. Annu. Rev. Med. 54, 17–28 (2003).

    Article  CAS  PubMed  Google Scholar 

  7. Zetter, B.R. Angiogenesis and tumor metastasis. Annu. Rev. Med. 49, 407–424 (1998).

    Article  CAS  PubMed  Google Scholar 

  8. Hata, K. et al. Expression of angiopoietin-1, angiopoietin-2, and Tie2 genes in normal ovary with corpus luteum and in ovarian cancer. Oncology 62, 340–348 (2002).

    Article  CAS  PubMed  Google Scholar 

  9. Tanaka, S. et al. Tie2 vascular endothelial receptor expression and function in hepatocellular carcinoma. Hepatology 35, 861–867 (2002).

    Article  CAS  PubMed  Google Scholar 

  10. Stratmann, A., Risau, W. & Plate, K.H. Cell type-specific expression of angiopoietin-1 and angiopoietin-2 suggests a role in glioblastoma angiogenesis. Am. J. Pathol. 153, 1459–1466 (1998).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Peters, K.G. et al. Expression of Tie2/Tek in breast tumour vasculature provides a new marker for evaluation of tumour angiogenesis. Br. J. Cancer 77, 51–56 (1998).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Lin, P. et al. Inhibition of tumor angiogenesis using a soluble receptor establishes a role for Tie2 in pathologic vascular growth. J. Clin. Invest. 100, 2072–2078 (1997).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Lin, P. et al. Antiangiogenic gene therapy targeting the endothelium specific receptor tyrosine kinase Tie2. Proc. Natl. Acad. Sci. USA 95, 8829–8834 (1998).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Ramsauer, M. & D'Amore, P.A. Getting Tie(2)d up in angiogenesis. J. Clin. Invest. 110, 1615–1617 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. 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  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  18. Fiedler, U. et al. Angiopoietin-1 and Angiopoietin-2 share the same binding domains in the Tie2 receptor involving the first Ig-like loop and the epidermal growth factor-like repeats. J. Biol. Chem. 278, 1721–1727 (2003).

    Article  CAS  PubMed  Google Scholar 

  19. Barton, W.A., Tzvetkova, D. & Nikolov, D.B. Structure of the Angiopoietin-2 receptor binding-domain and identification of surfaces involved in receptor recognition. Structure 13, 825–832 (2005).

    Article  CAS  PubMed  Google Scholar 

  20. Davis, S. et al. Angiopoietins have distinct modular domains essential for receptor binding, dimerization, and superclustering. Nat. Struct. Biol. 10, 38–44 (2003).

    Article  CAS  PubMed  Google Scholar 

  21. Ward, N.L. & Dumont, D.J. The angiopoietins and Tie2/Tek: adding to the complexity of cardiovascular development. Semin. Cell Dev. Biol. 13, 19–27 (2002).

    Article  CAS  PubMed  Google Scholar 

  22. Kim, K.-T. et al. Oligomerization and multimerization are critical for Angiopoietin-1 to bind and phosphorylate Tie2. J. Biol. Chem. 280, 20126–20131 (2005).

    Article  CAS  PubMed  Google Scholar 

  23. Wiesmann, C., Muller, Y.A. & de Vos, A.M. Ligand-binding sites in Ig-like domains of receptor tyrosine kinases. J. Mol. Med. 78, 247–260 (2000).

    Article  CAS  PubMed  Google Scholar 

  24. Hubbard, S.R. & Till, J.H. Protein tyrosine kinase structure and function. Annu. Rev. Biochem. 69, 373–379 (2000).

    Article  CAS  PubMed  Google Scholar 

  25. Dumont, D.J., Yamaguchi, T.P., Conlon, R.A., Rossant, J. & Breitman, M.L. tek, a novel tyrosine kinase gene located on mouse chromosome 4, is expressed in endothelial cells and their presumptive precursors. Oncogene 7, 1471–1480 (1992).

    CAS  PubMed  Google Scholar 

  26. Ziegler, S.F., Bird, T.A., Schneringer, J.A., Schooley, K.A. & Baum, P.R. Molecular cloning and characterization of a novel receptor protein tyrosine kinase from human placenta. Oncogene 8, 663–670 (1993).

    CAS  PubMed  Google Scholar 

  27. Harpaz, Y. & Chothia, C. Many of the immunoglobulin superfamily domains in cell adhesion molecules and surface receptors belong to a new structural set which is close to that containing variable domains. J. Mol. Biol. 238, 528–539 (1994).

    Article  CAS  PubMed  Google Scholar 

  28. Casasnovas, J.M., Stehle, T., Liu, J.H., Wang, J.H. & Springer, T.A. A dimeric crystal structure for the N-terminal two domains of intercellular adhesion molecule-1. Proc. Natl. Acad. Sci. USA 95, 4134–4139 (1998).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Freigang, J. et al. The crystal structure of the ligand binding module of axonin-1/TAG-1 suggests a zipper mechanism for neural cell adhesion. Cell 101, 425–433 (2000).

    Article  CAS  PubMed  Google Scholar 

  30. Zdanov, A. et al. Structure of a single-chain antibody variable domain (Fv) fragment complexed with a carbohydrate antigen at 1.7 Å resolution. Proc. Natl. Acad. Sci. USA 91, 6423 (1994).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Breithaupt, C. et al. Structural insights into the antigenicity of myelin oligodendrocyte glycoprotein. Proc. Natl. Acad. Sci. USA 100, 9446 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Yee, V.C. et al. Crystal structure of a 30 kDa C-terminal fragment from the γ chain of human fibrinogen. Structure 5, 125–138 (1997).

    Article  CAS  PubMed  Google Scholar 

  33. Pratt, K.P., Cote, H.C.F., Chung, D.W., Stenkamp, R.E. & Davie, E.W. The primary fibrin polymerization pocket: Three-dimensional structure of a 30 kDa C-terminal γ chain fragment complexed with the peptide Gly-Pro-Arg-Pro. Proc. Natl. Acad. Sci. USA 94, 7176–7181 (1997).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Collaborative Computational Project Number 4. The CCP4 suite: programs for X-ray crystallography. Acta Crystallogr. D Biol. Crystallogr. 50, 760–763 (1994).

  35. Braden, B.C. & Poljak, R.J. Structural features of the reactions between antibodies and protein antigens. FASEB J. 9, 9–16 (1995).

    Article  CAS  PubMed  Google Scholar 

  36. Sundberg, E.J. & Mariuzza, R.A. Molecular recognition in antibody-antigen complexes. Adv. Protein Chem. 61, 119–160 (2002).

    Article  PubMed  Google Scholar 

  37. Kairies, N. et al. The 2.0 Å crystal structure of tachylectin 5A provides evidence for the common origin of the innate immunity and the blood coagulation systems. Proc. Natl. Acad. Sci. USA 98, 13519–13524 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Stein, E. et al. Eph receptors discriminate specific ligand oligomers to determine alternative signaling complexes, attachment, and assembly responses. Genes Dev. 12, 667–678 (1998).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. 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  PubMed  Google Scholar 

  40. Carlson, T.R., Feng, Y., Maisonpierre, P.C., Mrksich, M. & Morla, A.O. Direct cell adhesion to the angiopoietins mediated by integrins. J. Biol. Chem. 276, 26516–26525 (2005).

    Article  Google Scholar 

  41. Wiesmann, C. et al. Crystal structure at 1.7Å resolution of VEGF in complex with domain 2 of the Flt-1 receptor. Cell 91, 695–704 (1997).

    Article  CAS  PubMed  Google Scholar 

  42. Ogiso, H. et al. Crystal structure of the complex of human epidermal growth factor and receptor extracellular domains. Cell 110, 775–787 (2002).

    Article  CAS  PubMed  Google Scholar 

  43. Garrett, T.P. et al. Crystal structure of a truncated epidermal growth factor receptor extracellular domain bound to transforming growth factor alpha. Cell 110, 763–773 (2002).

    Article  CAS  PubMed  Google Scholar 

  44. Himanen, J.P. & Nikolov, D.B. Eph signaling: a structural view. Trends Neurosci. 26, 46–51 (2003).

    Article  CAS  PubMed  Google Scholar 

  45. Otwinowski, Z. & Minor, W. Processing of X-ray diffraction data collected in oscillation mode. Methods Enzymol. 276, 307–326 (1997).

    Article  CAS  PubMed  Google Scholar 

  46. Weeks, C.M. & Miller, R. The design and implementation of SnB v2.0. J. Appl. Crystallogr. 32, 120–124 (1999).

    Article  CAS  Google Scholar 

  47. Evans, G. & Bricogne, G. Triiodide derivatization and combinatorial counter-ion replacement: two methods for enhancing phasing signal using laboratory Cu Kalpha X-ray equipment. Acta Crystallogr. D Biol. Crystallogr. 58, 976–991 (2002).

    Article  PubMed  Google Scholar 

  48. Jones, T.A., Zou, J.Y., Cowan, S.W. & Kjeldgaard, M. Improved methods for building protein models in electron density maps and the location of errors in these models. Acta Crystallogr. A 47, 110–119 (1991).

    Article  PubMed  Google Scholar 

  49. Brunger, A.T. et al. Crystallography and NMR system (CNS): A new software system for macromolecular structure determination. Acta Crystallogr. D Biol. Crystallogr. 54, 905–921 (1998).

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by the US National Institutes of Health (RO1-HL077249). We thank the staffs of beamlines X29A at the NSLS and NE-CAT at the Advanced Photon Source. Operation of the NSLS synchrotron beamline is supported by the US Department of Energy (contract no. DE-AC02-98CH10886). NE-CAT synchrotron beamline operations are supported by the National Center for Research Resources at the US National Institutes of Health (award RR-15301).

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Author notes

  1. William A Barton

    Present address: Department of Biochemistry, Virginia Commonwealth University, Richmond, Virginia, 23298, USA

  2. Kanagalaghatta R Rajashankar

    Present address: NE-CAT, Advanced Photon Source, Argonne, Illinois, 60439, USA

  3. William A Barton and Dorothea Tzvetkova-Robev: These authors contributed equally to this work.

Authors and Affiliations

  1. Structural Biology Program, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, 10021, New York, USA

    William A Barton, Edward P Miranda, Momchil V Kolev, Juha P Himanen & Dimitar B Nikolov

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Supplementary Fig. 1

A representative region of the experimental SAD electron density (PDF 1288 kb)

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Barton, W., Tzvetkova-Robev, D., Miranda, E. et al. Crystal structures of the Tie2 receptor ectodomain and the angiopoietin-2–Tie2 complex. Nat Struct Mol Biol 13, 524–532 (2006). https://doi.org/10.1038/nsmb1101

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