The measurement of the rotational diffusion coefficient of bovine plasma fibronectin by electric birefringence technique

Abstract

Although the conformation of fibronectin has been widely investigated by various techniques, there has not yet been any determination of its rotational diffusion coefficient. We report here this determination by the transient electric birefringence study of solutions of bovine plasma fibronectin at physiological ionic strength. The solutions showed a positive birefringence. A linear relationship was observed between the intensity of the birefringence at equilibrium and the square of the electric field within the range of fields applied (up to 12.5 kV.cm-1). The field-independent decay of the induced birefringence was described by a single exponential with a relaxation time of 0.76 (+/- 0.08) microsecond at 23 degrees C. This establishes fibronectin in solution as a globally rigid structure with a rotational diffusion coefficient, at 20 degrees C, of 202,000 s-1. This result allows the first rigorous determination of the low-resolution structure of fibronectin. It is important to notice that the analysis combines only results obtained in physiological conditions on native molecules and follows a strict hydrodynamic interpretation. The conclusion of this work is that a hollow sphere of about 20 nm external diameter can be proposed as a model for the three-dimensional structure of the fibronectin molecule in solution. This new model suggests the fibronectin could have the structure of a carrier protein.