Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Structure of a heparin-linked biologically active dimer of fibroblast growth factor

Nature volume 393, pages 812–817 (1998)Cite this article

Abstract

The fibroblast growth factors (FGFs) form a large family of structurally related, multifunctional proteins that regulate various biological responses1. They mediate cellular functions by binding to transmembrane FGF receptors2, which are protein tyrosine kinases. FGF receptors are activated by oligomerization3, and both this activation and FGF-stimulated biological responses require heparin-like molecules as well as FGF4. Heparins are linear anionic polysaccharide chains; they are typically heterogeneously sulphated on alternating L-iduronic and D-glucosamino sugars, and are nearly ubiquitous in animal tissues as heparan sulphate proteoglycans on cell surfaces and in the extracellular matrix. Although several crystal structures have been described for FGF molecules in complexes with heparin-like sugars5,6,7, the nature of a biologically active complex has been unknown until now. Here we describe the X-ray crystal structure, at 2.9 Å resolution, of a biologically active dimer of human acidic FGF in a complex with a fully sulphated, homogeneous heparin decassacharide. The dimerization of heparin-linked acidic FGF observed here is an elegant mechanism for the modulation of signalling through combinatorial homodimerization and heterodimerization of the 12 known members of the FGF family.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Biological activity of aFGF–heparin and aFGF–decasaccharide complexes.
Figure 2: Structures of heparin-linked aFGF dimers.
Figure 3: a–d, Details of hydrogen bonding between aFGF protomers (ribbons) and heparin (ball-and-stick diagrams) in aFGF dimers.

Similar content being viewed by others

References

  1. Basilico, C. & Moscatelli, D. The FGF family of growth factors and oncogenes. Adv. Cancer Res. 59, 115–165 (1992).

    Article  CAS  Google Scholar 

  2. Jaye, M., Schlessinger, J. & Dionne, C. A. Fibroblast growth factor receptor tyrosine kinases: molecular analysis and signal transduction. Biochem. Biophys. Acta 1135, 185–199 (1992).

    Article  CAS  Google Scholar 

  3. Schlessinger, J. Signal transduction by allosteric receptor oligomerization. Trends Biochem. Sci. 13, 443–447 (1988).

    Article  CAS  Google Scholar 

  4. Spivak-Kroizman, T. et al. Heparin-induced oligomerization of FGF molecules is responsible for FGF receptor dimerization, activation, and cell proliferation. Cell 79, 1015–1024 (1994).

    Article  CAS  Google Scholar 

  5. Zhu, X., Hsu, B. T. & Rees, D. C. Structural studies of the binding of the anti-ulcer drug sucrose octasulfate to acidic fibroblast growth factor. Structure 1, 27–34 (1993).

    Article  CAS  Google Scholar 

  6. Ornitz, D. M. et al. FGF binding and FGF receptor activation by synthetic heparan-derived di- and trisaccharides. Science 268, 432–436 (1995).

    Article  ADS  CAS  Google Scholar 

  7. Faham, S., Hileman, R. E., Fromm, J. R., Linhardt, R. J. & Rees, D. C. Heparin structure and interactions with basic fibroblast growth factor. Science 271, 1116–1120 (1996).

    Article  ADS  CAS  Google Scholar 

  8. Mulloy, B., Forster, M. J., Jones, C. & Davies, D. B. NMR and molecular-modelling studies of the solution conformation of heparin. Biochem. J. 293, 849–858 (1993).

    Article  CAS  Google Scholar 

  9. Burgess, W. H., Shaheen, A. M., Hampton, B., Donohue, P. J. & Winkles, J. A. Structure-function studies of heparin-binding (acidic fibroblast) growth factor-1 using site-directed mutagenesis. J. Cell Biol. 45, 131–138 (1991).

    CAS  Google Scholar 

  10. Thompson, L. D., Pantoliano, M. W. & Springer, B. A. Energetic characterization of the basic fibroblast growth factor-heparin interaction: identification of the heparin binding domain. Biochemistry 33, 3831–3840 (1994).

    Article  CAS  Google Scholar 

  11. Ornitz, D. M., et al. Heparin is required for cell-free binding of basic fibroblast growth factor to a soluble receptor and for mitogenesis in whole cells. Mol. Cell. Biol. 12, 240–247 (1992).

    Article  CAS  Google Scholar 

  12. Moy, F. J. et al. Properly oriented heparin-decasaccharide-induced dimers are the biologically active form of basic fibroblast growth factor. Biochemistry 36, 4782–4791 (1997).

    Article  CAS  Google Scholar 

  13. Herr, A. B., Ornitz, D. M., Sasisekharan, R., Venkataraman, G. & Waksman, G. Heparin-induced self-association of fibroblast growth factor-2: evidence for two oligomerization processes. J. Biol. Chem. 272, 16382–16389 (1997).

    Article  CAS  Google Scholar 

  14. Springer, B. A. et al. Identification and concerted function of two receptor binding surfaces on basic fibroblast growth factor required for mitogenesis. J. Biol. Chem. 269, 26879–26884 (1994).

    CAS  PubMed  Google Scholar 

  15. Kan, M. et al. An essential heparin-binding domain in the fibroblast growth factor receptor kinase. Science 259, 1918–1921 (1993).

    Article  ADS  CAS  Google Scholar 

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

    Article  Google Scholar 

  17. van Holde, K. E. & Weischet, W. O. Boundary analysis of sedimentation-velocity experiments with monodisperse and paucidisperse solutes. Biopolymers 17, 1387–1403 (1978).

    Article  CAS  Google Scholar 

  18. Huang, J., Mohammadi, M., Rodrigues, G. A. & Schlessingerr, J. Reduced activation of RAF-1 and MAP kinase by a fibroblast growth factor receptor mutant deficient in stimulation of phosphotidylinositol hydrolysis. J. Biol. Chem. 270, 5065–5072 (1995).

    Article  CAS  Google Scholar 

  19. Jaye, M. et al. Human endothelial cell growth factor: cloning, nucleotide sequence, and chromosome localization. Science 233, 541–545 (1986).

    Article  ADS  CAS  Google Scholar 

  20. Hendrickson, W. A., Horton, J. R. & LeMaster, D. M. Selenomethionyl proteins produced for analysis by multiwavelength anomalous diffraction (MAD): a vehicle for direct determination of three-dimensional structure. EMBO J. 9, 1665–1672 (1990).

    Article  CAS  Google Scholar 

  21. Otwinowski, Z. DENZO 1-56-62(SERC Daresbury Laboratory, Warrington, UK, 1993).

    Google Scholar 

  22. Collaborative Computational Project no. 4. The CCP4 suite: programs for protein crystallography. Acta Crystallogr. D 50, 760–763 (1994).

  23. Navaza, J. AMoRe: an automated package for molecular replacement. Acta Crystallogr. A 50, 157–163 (1994).

    Article  Google Scholar 

  24. Hendrickson, W. A. Determination of macromolecular structures from anomalous diffraction of synchrotron radiation. Science 254, 51–58 (1991).

    Article  ADS  CAS  Google Scholar 

  25. Brünger, A. T. X-PLOR Version 3.1, A System for X-ray Crystallography and NMR(Yale University Press, New Haven and London, 1992).

    Google Scholar 

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

    Article  Google Scholar 

  27. Laskowski, R. A., MacArthur, M. W., Moss, D. S. & Thornton, J. M. PROCHECK: a program to check stereochemical quality of protein structures. J. Appl. Crystallogr. 26, 283–291 (1993).

    Article  CAS  Google Scholar 

  28. Evans, S. V. SETOR: hardware lighted three-dimensional solid model representations of macromolecules. J. Mol. Graph. 11, 134–138 (1993).

    Article  CAS  Google Scholar 

  29. Nicholls, A., Sharp, K. A. & Honig, B. Protein folding and association: insights from the interfacial and thermodynamic properties of hydrocarbons. Proteins 11, 281–296 (1991).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank C. Bingman, M. Cuff, L. Shapiro, H. Yamaguchi and C. Ogata for help in data collection; J. Arnez, K. Drickamer, B. Honig, S. Hubbard, J. Passner, M. Quesenberry, L. Shapiro, and H. Yamaguchi for discussions; P. Östergaard (Novo Nordisk) for heparin oligosaccharides; M. Gawinowicz (HHMI Columbia) for amino-acid analysis; D. King for electrospray mass spectrometry; and C. Turgeon and J. Hansen for analytical ultracentrifugation. This work was supported in part by a grant from the NIH. Beamline X4A at the National Synchrotron Light Source, a DOE faciity, is supported by the HHMI.

Author information

Authors and Affiliations

  1. Department of Biochemistry, and Molecular Biophysics, Columbia University, New York, 10032, New York, USA

    Anna D. DiGabriele, Denise I. Chen & Wayne A. Hendrickson

  2. Howard Hughes Medical Institute, Columbia University, New York, 10032, New York, USA

    Wayne A. Hendrickson

  3. Department of Pharmacology, New York University Medical Center, New York, 10016, New York, USA

    Irit Lax & Joseph Schlessinger

  4. Pharmacia & Upjohn, Lindhagensgatan 133, S-11287, Stockholm, Sweden

    Carl M. Svahn

  5. Rhne-Poulenc-Rorer, 500 Arcola Road, Collegeville, 19426, Pennsylvania, USA

    Michael Jaye

Authors
  1. Anna D. DiGabriele
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Irit Lax
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Denise I. Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Carl M. Svahn
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Michael Jaye
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Joseph Schlessinger
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Wayne A. Hendrickson
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Wayne A. Hendrickson.

Rights and permissions

Reprints and permissions

About this article

Cite this article

DiGabriele, A., Lax, I., Chen, D. et al. Structure of a heparin-linked biologically active dimer of fibroblast growth factor. Nature 393, 812–817 (1998). https://doi.org/10.1038/31741

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/31741

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing