Insights into the role of heparan sulphate in fibroblast growth factor signalling.

Signalling from the FGFs (fibroblast growth factors) is crucial for the correct development and homoeostasis of a wide range of cells and tissues. The FGF/FGFR (FGF receptor) signalling system forms an important paradigm for HS (heparan sulphate)-binding proteins, as both the growth factor and receptor bind to HS, and HS or heparin is an absolute requirement for full signalling. The FGF signalling system has been extremely well structurally characterized, and details of each interaction involved in forming a ternary complex of FGF-FGFR-heparin have been elucidated. Recent work has focused on a more thorough understanding of the nature of the FGF-heparin complex in particular, demonstrating that FGFs preferentially bind to similar sites on the co-receptor, and that FGF-FGFR pairs show greater specificity for heparin sulphation patterns than individual FGFs. Further work has suggested that FGF-FGFR-heparin signalling complexes contain one molecule of heparin only, and that when longer fragments of heparin are used to form FGF-FGFR-heparin complexes, multiple complexes form upon the saccharide. These observations form the basis of a model where the range of interactions that FGFs and FGFRs can form with one another and with HS may lead to the formation of complexes with more than two FGFR units. Therefore HS will be crucial to FGF signalling from the initial signalling event to the formation of large receptor clusters.

Sequence analyses and comparative modeling of fly and worm fibroblast growth factor receptors indicate that the determinants for FGF and heparin binding are retained in evolution.

The presence of a large number of fibroblast growth factors (FGFs) and multiple splice forms of their receptors (FGFRs) in higher vertebrates makes the three-dimensional (3D) analysis of FGF interactions with their receptors a formidable task. The situation differs in Caenorhabditis elegans (worm) and Drosophila melanogaster (fruit fly), where only one or two FGF and FGFR sequences have been identified. Structural studies of the FGF-FGFR complexes in such primitive organisms should reveal the basic features of the ligand-receptor interactions as they first emerged through evolution. We have analysed the sequences of worm and fly FGFs and FGFRs and used the recently determined crystal structure of the human FGF1-FGFR2-heparin ternary complex [Pellegrini, L., Burke, D.F., von Delft, F., Mulloy, B. and Blundell, T.L. (2000) Nature 407, 1029-34] to construct 3D models of the homologous complexes. In spite of a low sequence similarity with their human counterparts, key structural features required for ligand-receptor and protein-heparin binding in humans are conserved in the fly and worm FGF-FGFR-heparin complexes. Analyses of the models show that tertiary interactions that are not conserved in sequence are maintained through novel interactions or complementary mutations in the fly and worm sequences. The overall charge distributions observed in the human FGF-FGFR-heparin complex are retained in the fly and worm models. The arginine residue at position 253 in the linker region between the Ig-like domains D2 and D3 in the wild type fly and worm sequences is particularly striking, as the Pro253Arg mutation in humans is responsible for Apert syndrome. This change may enhance the affinity of receptors for their FGF molecules as observed in Apert mutants.