| Abstract: |
Uncontrolled fibroblast growth factor (FGF) signaling can lead to human diseases, necessitating multiple layers of self-regulatory control mechanisms to keep its activity in check. Herein, we demonstrate that FGF9 and FGF20 ligands undergo a reversible homodimerization, occluding their key receptor binding sites. To test the role of dimerization in ligand autoinhibition, we introduced structure-based mutations into the dimer interfaces of FGF9 and FGF20. The mutations weakened the ability of the ligands to dimerize, effectively increasing the concentrations of monomeric ligands capable of binding and activating their cognate FGF receptor in vitro and in living cells. Interestingly, the monomeric ligands exhibit reduced heparin binding, resulting in their increased radii of heparan sulfate-dependent diffusion and biologic action, as evidenced by the wider dilation area of ex vivo lung cultures in response to implanted mutant FGF9-loaded beads. Hence, our data demonstrate that homodimerization autoregulates FGF9 and FGF20's receptor binding and concentration gradients in the extracellular matrix. Our study is the first to implicate ligand dimerization as an autoregulatory mechanism for growth factor bioactivity and sets the stage for engineering modified FGF9 subfamily ligands, with desired activity for use in both basic and translational research. Copyright © 2009, American Society for Microbiology. All Rights Reserved. |
| Keywords: |
signal transduction; protein expression; unclassified drug; gene mutation; mutation; nonhuman; animal cell; mouse; animals; mice; animal tissue; cells, cultured; embryo; diffusion; mice, inbred c57bl; extracellular matrix; amino acid sequence; molecular sequence data; protein multimerization; ligands; pregnancy; binding site; models, molecular; dimerization; crystallography, x-ray; protein structure, quaternary; fibroblast growth factor; fibroblast growth factor 20; fibroblast growth factor 9; heparan sulfate; receptor binding; fibroblast growth factors; heparitin sulfate; receptor, fibroblast growth factor, type 1
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