Energetics of heparin binding to human acidic fibroblast growth factor.
The binding of low-molecular-weight heparin to an amino-terminal-truncated, 132-amino-acid, human acidic fibroblast growth factor form has been studied by isothermal titration calorimetry. This technique yields values for the enthalpy change and equilibrium constant, from which the Gibbs energy and entropy change are also calculated. Experiments in different buffers and pH values show that the protonic balance during the reaction is negligible. Experiments made at pH 7.0 with NaCl concentrations ranging from 0.20 to 0.60 M revealed changes in enthalpy and Gibbs energy in the range of -30- -17 and -27- -24 kJ x mol(-1), respectively. Isothermal titration calorimetry was also performed at different temperatures to obtain a value for the heat-capacity change at pH 7.0 and 0.4 M NaCl concentration of -96 J K- x mol(-1). A change in the length of heparin brought about no change in the thermodynamic parameters at 25 degrees C under the same experimental conditions. Changes upon ligand binding in the range of -50- -200 A2 in both polar and non-polar solvent-accessible surface areas were calculated from thermodynamic data by using different parametric equations taken from the literature. These values suggest a negligible overall conformational change in the protein when it binds to heparin and no formation of any protein-protein interface.
Guzmán-Casado
Mercedes
M
Departamento de Química-Física e Instituto de Biotecnología, Facultad de Ciencias, Campus de Fuentenueva, Universidad de Granada, 18071, Granada, Spain.
García-Mira
María M
MM
Sánchez-Ruiz
José M
JM
Giménez-Gallego
Guillermo
G
Parody-Morreale
Antonio
A
D002151:Calorimetry; D004305:Dose-Response Relationship, Drug; D016220:Fibroblast Growth Factor 1 / Q000737:chemistry*; D006493:Heparin / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008024:Ligands; D011485:Protein Binding; D011487:Protein Conformation; D012964:Sodium / Q000494:pharmacology; D013696:Temperature; D013816:Thermodynamics
D008024:Ligands; D016220:Fibroblast Growth Factor 1; D006493:Heparin; D012964:Sodium