Glu-96 of Basic Fibroblast Growth Factor Is Essential for High Affinity Receptor Binding

Zhu, Hengyi; Ramnarayan, Kal; Anchin, Jerry M.; Miao, Wendy Y.; Sereno, Arlene; Millman, Linda; Jie, Zheng; Balaji, V.; Wolff, Manfred E.

doi:10.1074/jbc.270.37.21869

Cited by 27 publications

(27 citation statements)

References 43 publications

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“…Rat mtFGF contains a substitution of glutamine-104 with an alanine residue, equivalent to the glutamine-96 mutation on human FGF-2. This mutation produces FGF-2 that binds to the lowaffinity HSPG sites with unchanged affinity but has diminished affinity for FGFR1 (50). Our own characterization confirmed that, although mtFGF-2 was retained by the heart (presumably by binding to HSPGs of the basement membrane and the extracellular matrix) and localized around cardiomyocytes (Fig.…”

Section: Discussionsupporting

confidence: 65%

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Acute protection of ischemic heart by FGF-2: involvement of FGF-2 receptors and protein kinase C

Jiang

Padua

et al. 2002

American Journal of Physiology-Heart and Circulatory Physiology

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We examined the effect of fibroblast growth factor (FGF)-2 on myocardial resistance to injury when administered after the onset of ischemia , in vivo and ex vivo, and the role of FGF-2 receptors and protein kinase C (PKC). FGF-2 was injected into the left ventricle of rats undergoing permanent surgical coronary occlusion leading to myocardial infarction (MI). After 24 h, FGF-2-treated hearts displayed significantly reduced injury, determined by histological staining and troponin T release, and improved developed pressure compared with untreated controls. An FGF-2 mutant with diminished affinity for the tyrosine kinase FGF-2 receptor 1 (FGFR1) was not cardioprotective. FGF-2-treated hearts retained improved function and decreased damage at 6 wk after MI. In the ex vivo heart, FGF-2 administration during reperfusion after 30-min ischemia improved functional recovery and increased relative levels of PKC subtypes α, ε, and ζ in the particulate fraction, in a chelerythrine-preventable mode; it also decreased loss of energy metabolites. We conclude that intramyocardial FGF-2 administration shortly after the onset of ischemia confers protection from acute and chronic cardiac dysfunction and damage; FGF-2 delivered during reperfusion protects from ischemia-reperfusion injury; and protection by FGF-2 requires intact binding to FGFR1 and is likely mediated by PKC.

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Section: Discussionsupporting

confidence: 65%

“…2D) as well as irreversibly injured myocytes that have lost their antivinculin staining. No differences were evident in the staining pattern of wt-or mtFGF-2, as expected from their similar affinity to heparin (50).…”

Section: Resultsmentioning

confidence: 80%

Acute protection of ischemic heart by FGF-2: involvement of FGF-2 receptors and protein kinase C

Jiang

Padua

et al. 2002

American Journal of Physiology-Heart and Circulatory Physiology

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“…In conclusion, the data presented in this report show that FGF4 adopts a typical ␤-trefoil fold similar to other FGFs (24,28,43). A ternary FGF4-FGFR1-heparin model constructed by superimposing FGF4 onto FGF2 in the FGF2-FGFR1-heparin structure assisted the identification of several key residues in FGF4 involved in receptor and heparin binding.…”

Section: Resultsmentioning

confidence: 94%

“…This was particularly unexpected for the E159A mutant, because Glu-159 is highly conserved among FGFs (Fig. 1B) and the corresponding glutamic acid in FGF2 (Glu-96) was shown to be critical for binding of FGF2 to FGFR1 (43).…”

Section: Resultsmentioning

confidence: 99%

Identification of Receptor and Heparin Binding Sites in Fibroblast Growth Factor 4 by Structure-Based Mutagenesis

Bellosta¹,

Iwahori²,

Plotnikov

et al. 2001

Molecular and Cellular Biology

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Fibroblast growth factors (FGFs) comprise a large family of multifunctional, heparin-binding polypeptides that show diverse patterns of interaction with a family of receptors (FGFR1 to -4) that are subject to alternative splicing. FGFR binding specificity is an essential mechanism in the regulation of FGF signaling and is achieved through primary sequence differences among FGFs and FGFRs and through usage of two alternative exons, IIIc and IIIb, for the second half of immunoglobulin-like domain 3 (D3) in FGFRs. While FGF4 binds and activates the IIIc splice forms of FGFR1 to -3 at comparable levels, it shows little activity towards the IIIb splice forms of FGFR1 to -3 as well as towards FGFR4. To begin to explore the structural determinants for this differential affinity, we determined the crystal structure of FGF4 at a 1.8-Å resolution. FGF4 adopts a ␤-trefoil fold similar to other FGFs. To identify potential receptor and heparin binding sites in FGF4, a ternary FGF4-FGFR1-heparin model was constructed by superimposing the FGF4 structure onto FGF2 in the FGF2-FGFR1-heparin structure. Mutation of several key residues in FGF4, observed to interact with FGFR1 or with heparin in the model, produced ligands with reduced receptor binding and concomitant low mitogenic potential. Based on the modeling and mutational data, we propose that FGF4, like FGF2, but unlike FGF1, engages the ␤C-␤E loop in D3 and thus can differentiate between the IIIc and IIIb splice isoforms of FGFRs for binding. Moreover, we show that FGF4 needs to interact with both the 2-O-and 6-O-sulfates in heparin to exert its optimal biological activity.The fibroblast growth factor (FGF) family consists of 22 polypeptides (FGF1 to-22) with diverse biological activities (16,21,41). FGFs modulate proliferation and differentiation of a variety of cells of mesenchymal and neuroectodermal origin (1). FGFs play critical roles during embryonic processes such as mesoderm induction, postimplantation blastocyst development, and limb and lung development (7,40). Increased FGF signaling leads to a variety of human skeletal disorders, including dwarfism and craniosynostosis syndromes (15,19,39). In adult organisms, FGFs are thought to be involved in physiological angiogenesis and wound healing as well as in pathological angiogenesis, such as in tumor neovascularization and diabetic retinopathy (1).The diverse effects of FGFs are mediated by four receptor tyrosine kinases, FGFR1 to -4, which are composed of an extracellular ligand binding portion consisting of three immunoglobulin (Ig)-like domains (D1 to -3), a single transmembrane helix, and a cytoplasmic portion with protein tyrosine kinase activity. Ligand binding and specificity reside in D2, D3, and the short D2-D3 linker (29,30,34).Receptor dimerization is a prerequisite for FGF signaling and requires heparin or heparan sulfate proteoglycans (HSPGs) (22, 31). The recent crystal structure of a ternary FGF2-FGFR1-heparin complex has provided a mechanistic view of the process by which heparin aids FGFs to induce ...

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“…3. Five of the above seven amino acids (Y24, E96, N101, Y103, and L140) have been implicated to be important for receptor binding on the basis of site-directed mutagenesis studies (33,35). These amino acids form the primary FGFR binding site, as discussed below.…”

Section: Resultsmentioning

confidence: 99%

Molecular characteristics of fibroblast growth factor–fibroblast growth factor receptor–heparin-like glycosaminoglycan complex

Venkataraman

Raman

Sasisekharan

et al. 1999

Proc. Natl. Acad. Sci. U.S.A.

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Fibroblast growth factor (FGF) family plays key roles in development, wound healing, and angiogenesis. Understanding of the molecular nature of interactions of FGFs with their receptors (FGFRs) has been seriously limited by the absence of structural information on FGFR or FGF-FGFR complex. In this study, based on an exhaustive analysis of the primary sequences of the FGF family, we determined that the residues that constitute the primary receptor-binding site of FGF-2 are conserved throughout the FGF family, whereas those of the secondary receptor binding site of FGF-2 are not. We propose that the FGF-FGFR interaction mediated by the 'conserved' primary site interactions is likely to be similar if not identical for the entire FGF family, whereas the 'variable' secondary sites, on both FGF as well as FGFR mediates specificity of a given FGF to a given FGFR isoform. Furthermore, as the pro-inflammatory cytokine interleukin 1 (IL-1) and FGF-2 share the same structural scaffold, we find that the spatial orientation Fibroblast growth factors (FGFs) are important signaling molecules that play key roles during development and morphogenesis, as well as during several physiological and pathological situations such as wound healing, neovascularization, and tumor growth and progression (1-3). These growth factors induce mitogenic, chemotactic, and angiogenic activity in several different cell types (4). There are 18 FGF members in this important family, sharing between 10% and 55% sequence identity (5, 6). Despite only Ϸ55% sequence identity between FGF-1 and FGF-2, the nearly superimposable threedimensional structures of FGF-1 and FGF-2 suggest that all the members of the family share a common structural fold, namely the ␤-trefoil scaffold (7-9).FGFs signal though a set of four types of tyrosine kinase high-affinity receptors (FGFR-1 to -4) (10). The extracellular region of the FGFR contains three distinct Ig-like domains (I, II, and III). Alternative splicing of Ig III domain results in different spliced variants of each of the receptors (10). The different FGFRs exhibit unique affinities and selectivity for each of the FGF family members and the signal they transduce (11-13). In addition, the activity of these growth factors is also regulated by a low-affinity receptor, which has been identified to be heparin-like glycosaminoglycan (HLGAG), the polysaccharide component of the cell surface heparan sulfate proteoglycans (HSPGs) (3, 14). The macromolecular interactions of the growth factor, HLGAGs of HSPGs, and FGFR that lead to signal transduction is key to signaling by this important class of molecules (1,(14)(15)(16)(17). It has been suggested that HLGAGs mediate FGF dimerization or oligomerization, enabling the dimeric FGF to induce FGFR dimerization or oligomerization (18)(19)(20). The absence of structural information on the extracellular domains of the FGFRs has limited our understanding of the features and mechanisms of the interactions of FGF with FGFR and the role of HLGAGs in this process.Earlier, Pantol...

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Glu-96 of Basic Fibroblast Growth Factor Is Essential for High Affinity Receptor Binding

Cited by 27 publications

References 43 publications

Acute protection of ischemic heart by FGF-2: involvement of FGF-2 receptors and protein kinase C

Acute protection of ischemic heart by FGF-2: involvement of FGF-2 receptors and protein kinase C

Identification of Receptor and Heparin Binding Sites in Fibroblast Growth Factor 4 by Structure-Based Mutagenesis

Molecular characteristics of fibroblast growth factor–fibroblast growth factor receptor–heparin-like glycosaminoglycan complex

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