The structure of human acidic fibroblast growth factor and its interaction with heparin

Copeland, Robert A.; Ji, Hanlee P.; Halfpenny, Anne J.; Williams, Robert W.; Thompson, Karen; Herber, Wayne K.; Thomas, Kenneth A.; Bruner, Mark W.; Ryan, James A.; Marquis-Omer, Dorothy; Sanyal, Gautam; Sitrin, Robert D.; Yamazaki, Shigeko; Middaugh, C. Russell

doi:10.1016/0003-9861(91)90441-k

Cited by 124 publications

(137 citation statements)

References 40 publications

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“…FGF-1 is a weak mesophile as regards thermostability (DG unfolding ¼ 21.1 kJ/ mol), 25,28 and certain turn mutations can destabilize the protein by a magnitude approaching the overall DG unfolding ; thus, stabilizing background mutations were necessary for a subset of mutations. Of the 44 positions evaluated, 28 yielded |DDG| !…”

Section: Resultsmentioning

confidence: 99%

“…[37][38][39] Furthermore, the competent signal transduction complex of FGF-1 involves a ternary interaction between FGF-1, FGF receptor, and HSPG. [30][31][32][33] The addition of soluble heparin to FGF-1 confers resistance to thermal denaturation, chemical denaturation, and proteolysis, 28,40,41 and inclusion of heparin in the formulation of FGF-1 greatly improves its potency, stability, storage, and reconstitution properties. 28 Thus, heparin binding represents a key functionality that regulates the tissue distribution, pharmacokinetics, and receptor signaling of FGF-1.…”

Section: Discussionmentioning

confidence: 99%

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Experimental support for the foldability–function tradeoff hypothesis: Segregation of the folding nucleus and functional regions in fibroblast growth factor‐1

2012

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The acquisition of function is often associated with destabilizing mutations, giving rise to the stability-function tradeoff hypothesis. To test whether function is also accommodated at the expense of foldability, fibroblast growth factor-1 (FGF-1) was subjected to a comprehensive u-value analysis at each of the 11 turn regions. FGF-1, a b-trefoil fold, represents an excellent model system with which to evaluate the influence of function on foldability: because of its threefold symmetric structure, analysis of FGF-1 allows for direct comparisons between symmetryrelated regions of the protein that are associated with function to those that are not; thus, a structural basis for regions of foldability can potentially be identified. The resulting u-value distribution of FGF-1 is highly polarized, with the majority of positions described as either foldedlike or denatured-like in the folding transition state. Regions important for folding are shown to be asymmetrically distributed within the protein architecture; furthermore, regions associated with function (i.e., heparin-binding affinity and receptor-binding affinity) are localized to regions of the protein that fold after barrier crossing (late in the folding pathway). These results provide experimental support for the foldability-function tradeoff hypothesis in the evolution of FGF-1. Notably, the results identify the potential for folding redundancy in symmetric protein architecture with important implications for protein evolution and design.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Experimental support for the foldability–function tradeoff hypothesis: Segregation of the folding nucleus and functional regions in fibroblast growth factor‐1

2012

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“…One of the FGFR2c subunits in the ternary complex described by Pellegrini et al (27) was bound to an FGF1 molecule without additional heparin contacts. Heparin-HS interactions with FGF have previously been reported to reduce the conformational flexibility of the protein ligand (44,45). It is possible that binding to heparin saccharides might induce a subtle conformational change within FGF1 that stabilizes a favorable topology of the FGFR binding site.…”

Section: Discussionmentioning

confidence: 99%

Cooperative Dimerization of Fibroblast Growth Factor 1 (FGF1) upon a Single Heparin Saccharide May Drive the Formation of 2:2:1 FGF1·FGFR2c·Heparin Ternary Complexes

Robinson

Harmer

Goodger

et al. 2005

Journal of Biological Chemistry

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The related glycosaminoglycans heparin and heparan sulfate are essential for the activity of the fibroblast growth factor (FGF) family as they form an integral part of the signaling complex at the cell surface. Using size-exclusion chromatography we have studied the capacities of a variety of heparin oligosaccharides to bind FGF1 and FGFR2c both separately and together in ternary complexes. In the absence of heparin, FGF1 had no detectable affinity for FGFR2c. However, 2:2:1 complexes formed spontaneously in solution between FGF1, FGFR2c, and heparin octasaccharide (dp8). The dp8 sample was the shortest chain length that bound FGFR2c, that dimerized FGF1, and that promoted a strong mitogenic response to FGF1 through FGFR2c. Heparin hexasaccharide and various selectively desulfated heparin dp12s failed to bind FGFR2c and could only interact with FGF1 monomerically. These saccharides formed 1:1:1 complexes with FGF1 and FGFR2c, which had no tendency to self-associate, suggesting that binding of two FGF1 molecules to the same saccharide chain is a prerequisite for subsequent FGFR2c dimerization. We found that FGF1 dimerization upon heparin was favored over monomeric interactions even when a large excess of saccharide was present. A cooperative mechanism of FGF1 dimerization could explain how 2:2:1 signaling complexes form at the cell surface, an environment rich in heparan sulfate.The fibroblast growth factor (FGF) 2 family of proteins control the proliferation, migration, differentiation, and survival of a wide range of cell types (1). The FGFs are of fundamental importance during embryogenesis where they orchestrate the complex processes of organ formation and limb development (2). Dysregulated FGF expression is often associated with malignant transformation, because it facilitates the growth, survival, and metastatic spread of cancer cells (3). In humans 22 members of the FGF family have been described along with 4 FGF receptor tyrosine kinases (FGFR1-4). FGF signal transduction is achieved through ligand-dependent dimerization of FGFR subunits and their subsequent activation through transphosphorylation (4).The FGFRs share a number of common structural features (5). All have an intracellular split tyrosine kinase domain, a transmembrane sequence, and an extracellular region composed of three immunoglobulin (Ig)-like domains (D1-D3). FGF binding is mediated by the D2 and D3 domains, with the D3 domain having the primary influence on ligand specificity (6, 7). The D1 domain of the FGFRs does not take part in ligand interactions, and engineered receptors that lack this domain bind FGFs at least as strongly as full-length forms (8). For FGFRs1-3, alternative splicing of the D3 domain gives rise to two variants (termed IIIb and IIIc) with different ligand specificities (9). Therefore seven distinct FGF receptors can be expressed on the cell surface, each of which binds specifically to a subset of the FGFs. Significant redundancy is present within this system, because each receptor can be activated by several differe...

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“…FGF1 disrupts acidic pL-containing liposome integrity [29], is able to deform lipid bilayers [30], and it exhibits molten globule (MG) character at temperatures above 30°C [31,32], at acidic pH and in the presence of acidic pL [29]. MG is a partially unfolded conformation that bestows more hydrophobic features on the protein, and therefore increases the capability to interact with lipids [33].…”

Section: Introductionmentioning

confidence: 99%

Release of FGF1 and p40 synaptotagmin 1 correlates with their membrane destabilizing ability

Graziani

Bagalá

Duarte

et al. 2006

Biochemical and Biophysical Research Communications

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Fibroblast growth factor (FGF)1 is released from cells as a constituent of a complex that contains the small calcium binding protein S100A13, and the p40 kDa form of synaptotagmin (Syt)1, through an ER-Golgi-independent stress-induced pathway. FGF1 and the other components of its secretory complex are signal peptide-less proteins. We examined their capability to interact with lipid bilayers by studying protein-induced carboxyfluorescein release from liposomes of different phospholipid (pL) compositions. FGF1, p40 Syt1, and S100A13 induced destabilization of liposomes composed of acidic but not of zwitterionic pL. We produced mutants of FGF1 and p40 Syt1, in which specific basic amino acid residues in the regions that bind acidic pL were substituted. The ability of these mutants to induce liposomes destabilization was strongly attenuated, and they exhibited drastically diminished spontaneous and stress-induced release. Apparently, the non-classical release of FGF1 and p40 Syt1 involves destabilization of membranes containing acidic pL.

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The structure of human acidic fibroblast growth factor and its interaction with heparin

Cited by 124 publications

References 40 publications

Experimental support for the foldability–function tradeoff hypothesis: Segregation of the folding nucleus and functional regions in fibroblast growth factor‐1

Experimental support for the foldability–function tradeoff hypothesis: Segregation of the folding nucleus and functional regions in fibroblast growth factor‐1

Cooperative Dimerization of Fibroblast Growth Factor 1 (FGF1) upon a Single Heparin Saccharide May Drive the Formation of 2:2:1 FGF1·FGFR2c·Heparin Ternary Complexes

Release of FGF1 and p40 synaptotagmin 1 correlates with their membrane destabilizing ability

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