title: Thermodynamic analysis of the interaction of factor VIII with von Willebrand factor.
abstract: Factor VIII (FVIII) is a glycoprotein that plays an important role in the intrinsic pathway of coagulation. In circulation, FVIII is protected upon binding to von Willebrand factor (VWF), a chaperone molecule that regulates its half-life, distribution, and activity. Despite the biological significance of this interaction, its molecular mechanisms are not fully characterized. We determined the equilibrium and activation thermodynamics of the interaction between FVIII and VWF. The equilibrium affinity determined by surface plasmon resonance was temperature-dependent with a value of 0.8 nM at 35 °C. The FVIII-VWF interaction was characterized by very fast association (8.56 × 10(6) M(-1) s(-1)) and fast dissociation (6.89 × 10(-3) s(-1)) rates. Both the equilibrium association and association rate constants, but not the dissociation rate constant, were dependent on temperature. Binding of FVIII to VWF was characterized by favorable changes in the equilibrium and activation entropy (TΔS° = 89.4 kJ/mol, and -TΔS(++) = -8.9 kJ/mol) and unfavorable changes in the equilibrium and activation enthalpy (ΔH° = 39.1 kJ/mol, and ΔH(++) = 44.1 kJ/mol), yielding a negative change in the equilibrium Gibbs energy. Binding of FVIII to VWF in solid-phase assays demonstrated a high sensitivity to acidic pH and a sensitivity to ionic strength. Our data indicate that the interaction between FVIII and VWF is mediated mainly by electrostatic forces, and that it is not accompanied by entropic constraints, suggesting the absence of conformational adaptation but the presence of rigid "pre-optimized" binding surfaces.
authors.affiliation: Centre de Recherche des Cordeliers, Université Pierre et Marie Curie, Unité Mixte de Recherche S 872, Paris, France. jordan.dimitrov@crc.jussieu.fr
authors.forename: Jordan D
authors.initials: JD
authors.lastname: Dimitrov
authors.forename: Olivier D
authors.initials: OD
authors.lastname: Christophe
authors.forename: Jonghoon
authors.initials: J
authors.lastname: Kang
authors.forename: Yohann
authors.initials: Y
authors.lastname: Repessé
authors.forename: Sandrine
authors.initials: S
authors.lastname: Delignat
authors.forename: Srinivas V
authors.initials: SV
authors.lastname: Kaveri
authors.forename: Sébastien
authors.initials: S
authors.lastname: Lacroix-Desmazes
mesh_terms: D005169:Factor VIII / Q000737:chemistry*; D006801:Humans; D009994:Osmolar Concentration; D011485:Protein Binding; D055672:Static Electricity; D020349:Surface Plasmon Resonance; D013816:Thermodynamics; D014841:von Willebrand Factor / Q000737:chemistry*
chemical_list: D014841:von Willebrand Factor; D005169:Factor VIII