Solution NMR structure of a human FGF‐1 monomer, activated by a hexasaccharide heparin‐analogue

Canales, Ángeles; Lozano, Rosa M.; López-Méndez, Blanca; Angulo, Jesús; Ojeda, Ricardo A.; Nieto, Pedro M.; Martín‐Lomas, Manuel; Giménez‐Gallego, Guillermo; Jiménez‐Barbero, Jesús

doi:10.1111/j.1742-4658.2006.05474.x

Cited by 58 publications

(72 citation statements)

References 51 publications

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“…The use of oligosaccharides has already been validated as a good model to replace longer HS chains (44). Indeed, many structural and biological studies using oligosaccharides were consistent with the in vivo biological data (36,45). The minimal binding unit for CypB has previously been shown to be an octasaccharide (dp8).…”

Section: Discussionmentioning

confidence: 79%

Structural and Functional Characterization of the Interaction between Cyclophilin B and a Heparin-derived Oligosaccharide

Hanoulle

Melchior

Sibille

et al. 2007

Journal of Biological Chemistry

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The chemotaxis and integrin-mediated adhesion of T lymphocytes triggered by secreted cyclophilin B (CypB) depend on interactions with both cell surface heparan sulfate proteoglycans (HSPG) and the extracellular domain of the CD147 membrane receptor. Here, we use NMR spectroscopy to characterize the interaction of CypB with heparin-derived oligosaccharides. Chemical shift perturbation experiments allowed the precise definition of the heparan sulfate (HS) binding site of CypB. The N-terminal extremity of CypB, which contains a consensus sequence for heparin-binding proteins was modeled on the basis of our experimental NMR data. Because the HS binding site extends toward the CypB catalytic pocket, we measured its peptidyl-prolyl cis-trans isomerase (PPIase) activity in the absence or presence of a HS oligosaccharide toward a CD147-derived peptide. We report the first direct evidence that CypB is enzymatically active on CD147, as it is able to accelerate the cis/trans isomerization of the Asp 179 -Pro 180 bond in a CD147-derived peptide. However, HS binding has no significant influence on this PPIase activity. We thus conclude that the glycanic moiety of HSPG serves as anchor for CypB at the cell surface, and that the signal could be transduced by CypB via its PPIase activity toward CD147.First characterized as the molecular targets of the immunosuppressive drug cyclosporin A (CsA), 3 cyclophilins (Cyps) constitute one class of the prolyl cis/trans isomerases that catalyze the cis/trans interconversion of the peptide bond preceding a proline (1, 2). Members of this class such as the predominantly cytoplasmic CypA, the secreted CypB, and the mitochondrial CypD are small ubiquitous proteins sharing a high sequence homology (65% identity between human CypA and CypB), that translates into a closely related three-dimensional fold. Indeed, the NMR and crystal structures of CypA free and in complex with CsA (3-6), as well as the crystal structure of CypB in complex with a cyclosporine analogue (7) all show the same core structure composed of eight antiparallel ␤-strands forming a ␤-barrel surrounded by ␣-helices and loops. Whereas the nearly identical active site and CsA binding pocket further underscore their close relationship, both proteins do differ in their N and C termini, CypB containing two peptides of some 10 residues long that are lacking in CypA.CypA and CypB act in the progression of inflammatory diseases such as rheumatoid arthritis and psoriasis, but are equally involved in the first steps of certain viral infections (8 -10). Their inflammatory activity is conditioned by their interaction with heparan sulfate proteoglycans (HSPGs) and the membrane receptor CD147, two binding partners at the cell surface of T cell lymphocytes, granulocytes and macrophages (11-14). Significantly, both molecular partners have equally been described as co-receptors for the HIV-1 virus (10,12,15).Both intact prolyl cis/trans activity of the cyclophilins and the presence of the Pro 180 residue of CD147, located on one of the two ex...

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Section: Discussionmentioning

confidence: 79%

Structural and Functional Characterization of the Interaction between Cyclophilin B and a Heparin-derived Oligosaccharide

Hanoulle

Melchior

Sibille

et al. 2007

Journal of Biological Chemistry

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“…As the lower panel shows, there was a significant line broadening of most of the FGF-1 signals upon the addition of exdFGFR2IIIc, which causes many of them to remain hidden below the spectral noise. The framed crosspeaks corresponded to groups belonging to highly flexible regions of the protein, whose transverse relaxation time remained largely independent of the overall size of the polypeptide (35). Because the position and intensity of these crosspeaks remained practically the same in the upper and lower panels of the figure, the overall decrease in intensity of the rest of the signals cannot be attributed to 15 N-labeled FGF-1 coming out of solution upon the addition of exdFGFR2IIIc.…”

Section: 5dhps Competes With Heparin For Binding To Fgfrs-mentioning

confidence: 94%

“…The protocol to uniformly labeled 15 N synthesis has already been described (35). C-LYT/aFGF was produced and purified by heparinSepharose chromatography as described previously (36).…”

Section: Methodsmentioning

confidence: 99%

“…7B). The two-dimensional distribution of cross-peaks, already fully characterized, represented a fingerprint of the protein (35). The intensity of the signals in these spectra is inversely related to the size of the protein, as the line width of the signal is inversely proportional to the transverse relaxation time (T 2 ), which decreases as the efficiency of relaxation increases with the size of the molecule.…”

Section: 5dhps Competes With Heparin For Binding To Fgfrs-mentioning

confidence: 99%

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Gentisic Acid, a Compound Associated with Plant Defense and a Metabolite of Aspirin, Heads a New Class of in Vivo Fibroblast Growth Factor Inhibitors

Fernandez

Cuevas

Angulo

et al. 2010

Journal of Biological Chemistry

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106

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Fibroblast growth factors are key proteins in many intercellular signaling networks. They normally remain attached to the extracellular matrix, which confers on them a considerable stability. The unrestrained accumulation of fibroblast growth factors in the extracellular milieu, either due to uncontrolled synthesis or enzymatic release, contributes to the pathology of many diseases. Consequently, the neutralization of improperly mobilized fibroblast growth factors is of clear therapeutic interest. In pursuing described rules to identify potential inhibitors of these proteins, gentisic acid, a plant pest-controlling compound, an aspirin and vegetarian diet common catabolite, and a component of many traditional liquors and herbal remedies, was singled out as a powerful inhibitor of fibroblast growth factors. Gentisic acid was used as a lead to identify additional compounds with better inhibitory characteristics generating a new chemical class of fibroblast growth factor inhibitors that includes the agent responsible for alkaptonuria. Through low and high resolution approaches, using representative members of the fibroblast growth factor family and their cell receptors, it was shown that this class of inhibitors may employ two different mechanisms to interfere with the assembly of the signaling complexes that trigger fibroblast growth factor-driven mitogenesis. In addition, we obtained evidence from in vivo disease models that this group of inhibitors may be of interest to treat cancer and angiogenesis-dependent diseases.The fibroblast growth factors (FGFs) 4 constitute one of the largest families of polypeptide growth factors. There are 22FGFs in humans and mice that differ significantly in both size (17-20 kDa) and sequence, although each contains a core homology region encompassing 120 -130 residues. Phylogenetic analyses suggest that the FGF genes can be arranged into seven subfamilies. All FGFs bind to heparin with high affinity (K d between 1-2 nM), except for the members of the FGF-19 subfamily (i.e.: FGF-15, -19, -21, and -23) that have little or no affinity for these glycosaminoglycans (1). Apart from the family comprising FGF-11 to FGF-14, FGFs exert their diverse biological actions by binding to a series of membrane tyrosine kinase receptors (FGFRs) that are encoded by four genes (2-4). For this reason the FGF family is currently considered to be constituted by 18 members.FGFs were first isolated in the 1980s from bovine brain extracts due to their mitogenic and angiogenic activities (5). The affinity of FGFs for heparin was recognized very soon after their discovery (6), although the physiological substrate for FGF in normal conditions is heparan sulfate, a proteoglycan whose glycoside moiety is a glycosaminoglycan like heparin. Although initially conceived as FGF traps and protectors, it was later shown that these proteoglycans also participate in FGF signaling, although they are not absolutely required (4,7,8).In addition to the effects on cell replication and angiogenesis observed initially, FGFs r...

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“…Furthermore, it has been reported that the conformational equilibrium remains even when the hexasaccharide 2 is within the complex with FGF-1 as a result of interactions with flexible side chains via electrostatic charges. 52,53 The solution structure of the heparin was described by Mulloy et al as an extended helix with four residues per turn 20 that leads to clustering three consecutive sulphate groups directed towards the same side, relative to the central chain, while the subsequent cluster is directed towards the opposite side. The distribution of sulphate group charges in 1 can be considered to be analogous to that of compound 2 but in the reversed directionality (from the reducing to the non-reducing end; see Fig.…”

Section: Discussionmentioning

confidence: 99%

3D structure of a heparin mimetic analogue of a FGF-1 activator. A NMR and molecular modelling study

et al. 2013

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The motional behaviour of heparin oligosaccharides in solution is best described as a top rotor having two perpendicular rotation axes. This prevents an accurate extraction of interprotonic distances by NOESY/ROESY based methods. In this paper, we describe the solution structure of the hexasaccharide 1 calculated from high exactitude distance data obtained from off-resonance ROESY combined with a long MD simulation of 500 ns. In previous studies, we have found that two synthetic hexasaccharides having the sulphate groups directed towards one side of its central plane have an opposite biological activity, while 1 is unable to activate the FGF-1 signalling pathway, the other (2) is even more active than the regular region derived hexasaccharide (3) that mimics the natural active compound, heparin. From the structural analysis it was concluded that 1 has similar three-dimensional characteristics to 2 or 3 and therefore the differences in the activity should be due to the arrangement of the sulphate groups within the hexasaccharidic sequence.

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Solution NMR structure of a human FGF‐1 monomer, activated by a hexasaccharide heparin‐analogue

Cited by 58 publications

References 51 publications

Structural and Functional Characterization of the Interaction between Cyclophilin B and a Heparin-derived Oligosaccharide

Structural and Functional Characterization of the Interaction between Cyclophilin B and a Heparin-derived Oligosaccharide

Gentisic Acid, a Compound Associated with Plant Defense and a Metabolite of Aspirin, Heads a New Class of in Vivo Fibroblast Growth Factor Inhibitors

3D structure of a heparin mimetic analogue of a FGF-1 activator. A NMR and molecular modelling study

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