The intriguing diversity of published translational diffusion constants for the fibrinogen molecule can hardly be explained, unless interactions between the molecules are postulated. In the present study we have investigated the possible effect of molecular association and electrostatic intermolecular interactions on the Brownian motion. The translational diffusion coefficient DT, the rotational diffusion coefficient around the minor axis DR and the sedimentation coefficient have been measured. The methods used were dynamic light scattering and analytical ultracentrifugation. The samples were solutions of purified human fibrinogen. The correlation-function corresponding to DT deviates from a single exponential. The initial slope is found to depend on concentration, being DT = (1.7 ± 0.3) 10-7 cm2/s at 10mg/ml, pH 7.4 and 0.15 molar Tris-NaCl, and increases at fibrinogen concentrations below 2mg/ml. These results are compatible with a polydispers solution, in which single molecules are in equilibrium with pair and higher aggregates. The nature of the aggregates is end-to-end as indicated from the difference between the two rotational diffusion constants DR = 40000 ± 20% and DR = 10000 ±30% s-1. On the basis of the Hall-Slayter model and assumption of end-to-end association we calculated the ratio of the sedimentation coefficient of single, pair and triplet associates, being 1:1.14:1.20. Therefore, it is difficult to separate them in a sedimentation run. For ionic strength below 0.05 molar and low fibrinogen concentration (0.lmg/ml) a fast decay appears in the correlation, indicating that the Brownian motion is strongly influenced by electrostatic interactions.
Brownian motion: Theory and experiment a simple classroom measurement of the diffusion coefficient
