Myo-inositol hexasulphate and low molecular weight heparin binding to human acidic fibroblast growth factor: a calorimetric and FTIR study

Guzmán-Casado, Mercedes; Cardenete, A.; Giménez‐Gallego, Guillermo; Parody-Morreale, Antonio

doi:10.1016/s0141-8130(01)00131-3

Cited by 13 publications

(7 citation statements)

References 33 publications

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“…Furthermore, the choice of buffer and pH can influence the heat capacity change (44); thus caution must be used in interpreting these data. However, the value of ΔC p (-644 J K -1 mol -1 ) is similar to values obtained for other proteins binding to heparin (52).…”

Section: Discussionsupporting

confidence: 84%

Thermodynamic Characterization of the Interaction between the C-Terminal Domain of Extracellular Superoxide Dismutase and Heparin by Isothermal Titration Calorimetry

2009

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Extracellular superoxide dismutase (ECSOD) interacts with heparin through its C-terminal domain. In this study we used isothermal titration calorimetry (ITC) to get detailed thermodynamic information about the interaction. We have shown that the interaction between ECSOD and intestinal mucosal heparin (M(w) 6000-30000 Da) is exothermic and driven by enthalpy at physiological salt concentration. However, the contribution from entropy is favorable for binding of small isolated heparin fragments. By studying different size-defined heparin fragments, we also concluded that a hexasaccharide moiety is sufficient for strong binding to ECSOD. The binding involves proton transfer from the buffer to the ECSOD-heparin complex, and the results indicate that the number of ionic interactions made between ECSOD and heparin upon binding varies from three to five for heparin and an octasaccharide fragment, respectively. Surprisingly and despite the many charges found on both the protein and the polysaccharide, our results indicate that the nonionic contribution to the binding is large. From the temperature dependence we have calculated the constant pressure heat capacity change (DeltaC(p)) of the interaction to -644 J K(-1) mol(-1) and -306 J K(-1) mol(-1) for heparin and an octasaccharide, respectively.

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Section: Discussionsupporting

confidence: 84%

Thermodynamic Characterization of the Interaction between the C-Terminal Domain of Extracellular Superoxide Dismutase and Heparin by Isothermal Titration Calorimetry

2009

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“…On binding FGF-1, there was reportedly a small change in protein tertiary structure, but little change in secondary structure, 24 while the interaction between FGF-2 and heparin did alter secondary structure. 25 Nevertheless, both types of change in protein structure caused an increase in thermal stability but, this may not necessarily have resulted in the assembly of an active signalling complex.…”

Section: Resultsmentioning

confidence: 95%

“…This calls into question the level of unique information that HS sequences contain and backsup earlier work in which structural redundancy in HS analogues was predicted and modeled. 28 FGF-1 is known to have a disordered structure, often referred to as "molten globular" comprising ~55% b-strands and ~20% turns, ~10% a-helix and ~15% un-ordered stretches, 29 and undergoes a small change in tertiary structure 24 when binding sulfated polysaccharides. The structural requirements of the associated anionic saccharide for signalling through FGF-1 or FGF-2 with FGFR1c, which were examined here, clearly showed considerable latitude.…”

Section: Discussionmentioning

confidence: 99%

Comparable stabilisation, structural changes and activities can be induced in FGF by a variety of HS and non-GAG analogues: implications for sequence-activity relationships

et al. 2010

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The activities of heparan sulfate (HS) and heparin do not correlate simply with sulfation levels or sequence. The alternative hypothesis, that appropriate charge and conformational characteristics for protein binding and activity can be provided by other sequences in heparan sulfate and, possibly, also in unrelated sulfated polysaccharides, is explored. Differential scanning fluorimetry was used to measure the thermostabilisation bestowed by modified heparin polysaccharides (proxies for heparan sulfate) on fibroblast growth factor-1 (FGF-1) and fibroblast growth factor-2 (FGF-2), prototypical heparan sulfate-binding proteins, revealing varied abilities and primary sequence-activity redundancy. The effect of substitution pattern on the heparin/heparan sulfate backbone was explored using principal component analysis of (13)C NMR chemical shift data for homogeneously modified heparin polysaccharides revealing complex conformational effects. No simple relationship emerged between these polysaccharides, with their distinct charge distributions and geometries, and their ability to signal. Other, structurally unrelated sulfated polysaccharides were also able to support signalling. These influenced FGF stabilisation in a similar manner to the HS analogues and provided analogous cell signalling activity. For FGF-1, but not FGF-2, signaling correlated strongly with protein stabilisation and circular dichroism spectroscopy demonstrated that some non-HS polysaccharides invoked comparable secondary structural changes to those induced by heparin. Active conformations can readily be found in several heparin derivatives, as well as among non-HS polysaccharides, which comprise unrelated primary sequences, confirming the hypothesis and implying that the level of unique information contained in HS sequences may be much lower than previously thought.

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“…Many methods have been used to study the interactions between heparins and proteins such as isothermal titration calorimetry [6][7][8][9][10][11], surface plasmon resonance spectrometry (SPR) [5,6,8,12,13], affinity chromatography (AC) [6,[8][9][10]14,15], nuclear magnetic resonance spectrometry [5,6,16], X-ray [5,10,17,18], mass spectrometry [19], circular dichroism [11,14], Fourier transform infrared spectrometry [7], polyacrylamide gel electrophoresis [5], fluorescence [20], equilibrium dialysis [9,21] and capillary electrophoresis (CE) [19,22,23]. Among these methods, SPR and AC are widely used, but they require immobilization of either of the involved substances to supporting material and this poses a problem of steric hindrance [24,25].…”

Section: Introductionmentioning

confidence: 99%

Capillary zone electrophoresis characterization of low molecular weight heparin binding to interleukin 2

Liang

et al. 2005

Journal of Pharmaceutical and Biomedical Analysis

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Myo-inositol hexasulphate and low molecular weight heparin binding to human acidic fibroblast growth factor: a calorimetric and FTIR study

Cited by 13 publications

References 33 publications

Thermodynamic Characterization of the Interaction between the C-Terminal Domain of Extracellular Superoxide Dismutase and Heparin by Isothermal Titration Calorimetry

Thermodynamic Characterization of the Interaction between the C-Terminal Domain of Extracellular Superoxide Dismutase and Heparin by Isothermal Titration Calorimetry

Comparable stabilisation, structural changes and activities can be induced in FGF by a variety of HS and non-GAG analogues: implications for sequence-activity relationships

Capillary zone electrophoresis characterization of low molecular weight heparin binding to interleukin 2

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