title: Heparin binding preference and structures in the fibroblast growth factor family parallel their evolutionary diversification.
abstract: The interaction of a large number of extracellular proteins with heparan sulfate (HS) regulates their transport and effector functions, but the degree of molecular specificity underlying protein-polysaccharide binding is still debated. The 15 paracrine fibroblast growth factors (FGFs) are one of the paradigms for this interaction. Here, we measure the binding preferences of six FGFs (FGF3, FGF4, FGF6, FGF10, FGF17, FGF20) for a library of modified heparins, representing structures in HS, and model glycosaminoglycans, using differential scanning fluorimetry. This is complemented by the identification of the lysine residues in the primary and secondary binding sites of the FGFs by a selective labelling approach. Pooling these data with previous sets provides good coverage of the FGF phylogenetic tree, deduced from amino acid sequence alignment. This demonstrates that the selectivity of the FGFs for binding structures in sulfated polysaccharides and the pattern of secondary binding sites on the surface of FGFs follow the phylogenetic relationship of the FGFs, and so are likely to be the result of the natural selection pressures that led to the expansion of the FGF family in the course of the evolution of more complex animal body plans.
authors.affiliation: Department of Biochemistry, Institute of Integrative Biology, University of Liverpool, Biosciences Building, Crown Street, Liverpool L69 7ZB, UK.
authors.forename: Yong
authors.identifier: http://orcid.org/0000-0002-5594-4434
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Biochemistry, Institute of Integrative Biology, University of Liverpool, Biosciences Building, Crown Street, Liverpool L69 7ZB, UK.
authors.forename: Changye
authors.identifier: http://orcid.org/0000-0001-8602-9629
authors.initials: C
authors.lastname: Sun
authors.affiliation: Department of Biochemistry, Institute of Integrative Biology, University of Liverpool, Biosciences Building, Crown Street, Liverpool L69 7ZB, UK.
authors.forename: Edwin A
authors.identifier: http://orcid.org/0000-0001-9365-5433
authors.initials: EA
authors.lastname: Yates
authors.affiliation: School of Pharmaceutical Science, Wenzhou Medical University, Chashan University Park, Wenzhou 325035, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Jiang
authors.affiliation: Department of Biochemistry, Institute of Integrative Biology, University of Liverpool, Biosciences Building, Crown Street, Liverpool L69 7ZB, UK.
authors.forename: Mark C
authors.initials: MC
authors.lastname: Wilkinson
authors.affiliation: Department of Biochemistry, Institute of Integrative Biology, University of Liverpool, Biosciences Building, Crown Street, Liverpool L69 7ZB, UK dgferning@liverpool.ac.uk.
authors.forename: David G
authors.identifier: http://orcid.org/0000-0003-4875-4293
authors.initials: DG
authors.lastname: Fernig
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D005346:Fibroblast Growth Factors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006025:Glycosaminoglycans / Q000737:chemistry / Q000378:metabolism; D006497:Heparan Sulfate / Q000031:analogs & derivatives / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D010802:Phylogeny; D011485:Protein Binding; D011487:Protein Conformation; D055550:Protein Stability; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013552:Swine
chemical_list: D005346:Fibroblast Growth Factors; D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D011994:Recombinant Proteins
keywords: fibroblast growth factor; glycosaminoglycan; heparan sulfate; heparin binding site; molecular recognition; specificity