Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1996 Jan 23;93(2):845–850. doi: 10.1073/pnas.93.2.845

Preferential self-association of basic fibroblast growth factor is stabilized by heparin during receptor dimerization and activation.

G Venkataraman 1, V Sasisekharan 1, A B Herr 1, D M Ornitz 1, G Waksman 1, C L Cooney 1, R Langer 1, R Sasisekharan 1
PMCID: PMC40145  PMID: 8570646

Abstract

Central to signaling by fibroblast growth factors (FGFs) is the oligomeric interaction of the growth factor and its high-affinity cell surface receptor, which is mediated by heparin-like polysaccharides. It has been proposed that the binding of heparin-like polysaccharides to FGF induces a conformational change in FGF, resulting in the formation of FGF dimers or oligomers, and this biologically active form is 'presented' to the FGF receptor for signal transduction. In this study, we show that monomeric basic FGF (FGF-2) preferentially self-associates and forms FGF-2 dimers and higher-order oligomers. As a consequence, FGF-2 monomers are oriented for binding to heparin-like polysaccharides. We also show that heparin-like polysaccharides can readily bind to self-associated FGF-2 without causing a conformational change in FGF-2 or disrupting the FGF-2 self-association, but that the bound polysaccharides only additionally stabilize the FGF-2 self-association. The preferential self-association corresponds to FGF-2 translations along two of the unit cell axes of the FGF-2 crystal structures. These two axes represent the two possible heparin binding directions, whereas the receptor binding sites are oriented along the third axis. Thus, we propose that preferential FGF-2 self-association, further stabilized by heparin, like "beads on a string," mediates FGF-2-induced receptor dimerization and activation. The observed FGF-2 self-association, modulated by heparin, not only provides a mechanism of growth factor activation but also represents a regulatory mechanism governing FGF-2 biological activity.

Full text

PDF
845

Images in this article

847Fig. 1
on p.847
847Fig. 2
on p.847
847Fig. 3
on p.847
848Fig. 4
on p.848
848Fig. 5
on p.848

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ago H., Kitagawa Y., Fujishima A., Matsuura Y., Katsube Y. Crystal structure of basic fibroblast growth factor at 1.6 A resolution. J Biochem. 1991 Sep;110(3):360–363. doi: 10.1093/oxfordjournals.jbchem.a123586. [DOI] [PubMed] [Google Scholar]
  2. Aviezer D., Hecht D., Safran M., Eisinger M., David G., Yayon A. Perlecan, basal lamina proteoglycan, promotes basic fibroblast growth factor-receptor binding, mitogenesis, and angiogenesis. Cell. 1994 Dec 16;79(6):1005–1013. doi: 10.1016/0092-8674(94)90031-0. [DOI] [PubMed] [Google Scholar]
  3. Baird A., Schubert D., Ling N., Guillemin R. Receptor- and heparin-binding domains of basic fibroblast growth factor. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2324–2328. doi: 10.1073/pnas.85.7.2324. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Basilico C., Moscatelli D. The FGF family of growth factors and oncogenes. Adv Cancer Res. 1992;59:115–165. doi: 10.1016/s0065-230x(08)60305-x. [DOI] [PubMed] [Google Scholar]
  5. Bernfield M., Kokenyesi R., Kato M., Hinkes M. T., Spring J., Gallo R. L., Lose E. J. Biology of the syndecans: a family of transmembrane heparan sulfate proteoglycans. Annu Rev Cell Biol. 1992;8:365–393. doi: 10.1146/annurev.cb.08.110192.002053. [DOI] [PubMed] [Google Scholar]
  6. Burgess W. H., Maciag T. The heparin-binding (fibroblast) growth factor family of proteins. Annu Rev Biochem. 1989;58:575–606. doi: 10.1146/annurev.bi.58.070189.003043. [DOI] [PubMed] [Google Scholar]
  7. Eriksson A. E., Cousens L. S., Matthews B. W. Refinement of the structure of human basic fibroblast growth factor at 1.6 A resolution and analysis of presumed heparin binding sites by selenate substitution. Protein Sci. 1993 Aug;2(8):1274–1284. doi: 10.1002/pro.5560020810. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Eriksson A. E., Cousens L. S., Weaver L. H., Matthews B. W. Three-dimensional structure of human basic fibroblast growth factor. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3441–3445. doi: 10.1073/pnas.88.8.3441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Folkman J., Klagsbrun M., Sasse J., Wadzinski M., Ingber D., Vlodavsky I. A heparin-binding angiogenic protein--basic fibroblast growth factor--is stored within basement membrane. Am J Pathol. 1988 Feb;130(2):393–400. [PMC free article] [PubMed] [Google Scholar]
  10. Givol D., Yayon A. Complexity of FGF receptors: genetic basis for structural diversity and functional specificity. FASEB J. 1992 Dec;6(15):3362–3369. [PubMed] [Google Scholar]
  11. Guimond S., Maccarana M., Olwin B. B., Lindahl U., Rapraeger A. C. Activating and inhibitory heparin sequences for FGF-2 (basic FGF). Distinct requirements for FGF-1, FGF-2, and FGF-4. J Biol Chem. 1993 Nov 15;268(32):23906–23914. [PubMed] [Google Scholar]
  12. Heldin C. H. Dimerization of cell surface receptors in signal transduction. Cell. 1995 Jan 27;80(2):213–223. doi: 10.1016/0092-8674(95)90404-2. [DOI] [PubMed] [Google Scholar]
  13. Ishihara M., Tyrrell D. J., Stauber G. B., Brown S., Cousens L. S., Stack R. J. Preparation of affinity-fractionated, heparin-derived oligosaccharides and their effects on selected biological activities mediated by basic fibroblast growth factor. J Biol Chem. 1993 Mar 5;268(7):4675–4683. [PubMed] [Google Scholar]
  14. Janin J., Miller S., Chothia C. Surface, subunit interfaces and interior of oligomeric proteins. J Mol Biol. 1988 Nov 5;204(1):155–164. doi: 10.1016/0022-2836(88)90606-7. [DOI] [PubMed] [Google Scholar]
  15. Klagsbrun M., Baird A. A dual receptor system is required for basic fibroblast growth factor activity. Cell. 1991 Oct 18;67(2):229–231. doi: 10.1016/0092-8674(91)90173-v. [DOI] [PubMed] [Google Scholar]
  16. Maccarana M., Casu B., Lindahl U. Minimal sequence in heparin/heparan sulfate required for binding of basic fibroblast growth factor. J Biol Chem. 1993 Nov 15;268(32):23898–23905. [PubMed] [Google Scholar]
  17. Mason I. J. The ins and outs of fibroblast growth factors. Cell. 1994 Aug 26;78(4):547–552. doi: 10.1016/0092-8674(94)90520-7. [DOI] [PubMed] [Google Scholar]
  18. McDonald N. Q., Lapatto R., Murray-Rust J., Gunning J., Wlodawer A., Blundell T. L. New protein fold revealed by a 2.3-A resolution crystal structure of nerve growth factor. Nature. 1991 Dec 5;354(6352):411–414. doi: 10.1038/354411a0. [DOI] [PubMed] [Google Scholar]
  19. Mulloy B., Forster M. J., Jones C., Davies D. B. N.m.r. and molecular-modelling studies of the solution conformation of heparin. Biochem J. 1993 Aug 1;293(Pt 3):849–858. doi: 10.1042/bj2930849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nugent M. A., Edelman E. R. Kinetics of basic fibroblast growth factor binding to its receptor and heparan sulfate proteoglycan: a mechanism for cooperactivity. Biochemistry. 1992 Sep 22;31(37):8876–8883. doi: 10.1021/bi00152a026. [DOI] [PubMed] [Google Scholar]
  21. Ornitz D. M., Herr A. B., Nilsson M., Westman J., Svahn C. M., Waksman G. FGF binding and FGF receptor activation by synthetic heparan-derived di- and trisaccharides. Science. 1995 Apr 21;268(5209):432–436. doi: 10.1126/science.7536345. [DOI] [PubMed] [Google Scholar]
  22. Ornitz D. M., Yayon A., Flanagan J. G., Svahn C. M., Levi E., Leder P. 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. 1992 Jan;12(1):240–247. doi: 10.1128/mcb.12.1.240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pantoliano M. W., Horlick R. A., Springer B. A., Van Dyk D. E., Tobery T., Wetmore D. R., Lear J. D., Nahapetian A. T., Bradley J. D., Sisk W. P. Multivalent ligand-receptor binding interactions in the fibroblast growth factor system produce a cooperative growth factor and heparin mechanism for receptor dimerization. Biochemistry. 1994 Aug 30;33(34):10229–10248. doi: 10.1021/bi00200a003. [DOI] [PubMed] [Google Scholar]
  24. Rapraeger A. C., Krufka A., Olwin B. B. Requirement of heparan sulfate for bFGF-mediated fibroblast growth and myoblast differentiation. Science. 1991 Jun 21;252(5013):1705–1708. doi: 10.1126/science.1646484. [DOI] [PubMed] [Google Scholar]
  25. Rapraeger A. C. The coordinated regulation of heparan sulfate, syndecans and cell behavior. Curr Opin Cell Biol. 1993 Oct;5(5):844–853. doi: 10.1016/0955-0674(93)90034-n. [DOI] [PubMed] [Google Scholar]
  26. Roghani M., Mansukhani A., Dell'Era P., Bellosta P., Basilico C., Rifkin D. B., Moscatelli D. Heparin increases the affinity of basic fibroblast growth factor for its receptor but is not required for binding. J Biol Chem. 1994 Feb 11;269(6):3976–3984. [PubMed] [Google Scholar]
  27. Sasisekharan R., Moses M. A., Nugent M. A., Cooney C. L., Langer R. Heparinase inhibits neovascularization. Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1524–1528. doi: 10.1073/pnas.91.4.1524. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Spivak-Kroizman T., Lemmon M. A., Dikic I., Ladbury J. E., Pinchasi D., Huang J., Jaye M., Crumley G., Schlessinger J., Lax I. Heparin-induced oligomerization of FGF molecules is responsible for FGF receptor dimerization, activation, and cell proliferation. Cell. 1994 Dec 16;79(6):1015–1024. doi: 10.1016/0092-8674(94)90032-9. [DOI] [PubMed] [Google Scholar]
  29. Springer B. A., Pantoliano M. W., Barbera F. A., Gunyuzlu P. L., Thompson L. D., Herblin W. F., Rosenfeld S. A., Book G. W. Identification and concerted function of two receptor binding surfaces on basic fibroblast growth factor required for mitogenesis. J Biol Chem. 1994 Oct 28;269(43):26879–26884. [PubMed] [Google Scholar]
  30. Tainer J. A., Getzoff E. D., Richardson J. S., Richardson D. C. Structure and mechanism of copper, zinc superoxide dismutase. Nature. 1983 Nov 17;306(5940):284–287. doi: 10.1038/306284a0. [DOI] [PubMed] [Google Scholar]
  31. Vlodavsky I., Fuks Z., Ishai-Michaeli R., Bashkin P., Levi E., Korner G., Bar-Shavit R., Klagsbrun M. Extracellular matrix-resident basic fibroblast growth factor: implication for the control of angiogenesis. J Cell Biochem. 1991 Feb;45(2):167–176. doi: 10.1002/jcb.240450208. [DOI] [PubMed] [Google Scholar]
  32. Wells J. A. Structural and functional basis for hormone binding and receptor oligomerization. Curr Opin Cell Biol. 1994 Apr;6(2):163–173. doi: 10.1016/0955-0674(94)90132-5. [DOI] [PubMed] [Google Scholar]
  33. Yayon A., Klagsbrun M., Esko J. D., Leder P., Ornitz D. M. Cell surface, heparin-like molecules are required for binding of basic fibroblast growth factor to its high affinity receptor. Cell. 1991 Feb 22;64(4):841–848. doi: 10.1016/0092-8674(91)90512-w. [DOI] [PubMed] [Google Scholar]
  34. Zhang J. D., Cousens L. S., Barr P. J., Sprang S. R. Three-dimensional structure of human basic fibroblast growth factor, a structural homolog of interleukin 1 beta. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3446–3450. doi: 10.1073/pnas.88.8.3446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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. 1993 Sep 15;1(1):27–34. doi: 10.1016/0969-2126(93)90006-3. [DOI] [PubMed] [Google Scholar]
  36. Zhu X., Komiya H., Chirino A., Faham S., Fox G. M., Arakawa T., Hsu B. T., Rees D. C. Three-dimensional structures of acidic and basic fibroblast growth factors. Science. 1991 Jan 4;251(4989):90–93. doi: 10.1126/science.1702556. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES