Structure of fibrinogen in electrolyte solutions derived from dynamic light scattering (DLS) and viscosity measurements.
Bulk physicochemical properties of bovine plasma fibrinogen (Fb) in electrolyte solutions were characterized. These comprised determination of the diffusion coefficient (hydrodynamic radius), electrophoretic mobility, and isoelectric point (iep). The hydrodynamic radius of Fb for the ionic strength of 0.15 M was 12.7 nm for pH 7.4 (physiological conditions) and 12 nm for pH 9.5. Using these values, the number of uncompensated (electrokinetic) charges on the protein N(c) was calculated from the electrophoretic mobility data. It was found that for physiological condition (pH 7.4, I = 0.15), N(c) = -7.6. For pH 9.5 and I = 10(-2), N(c) = -26. On the other hand, N(c) became zero independent of the ionic strength at pH 5.8, which was identified as the iep. Consequently, for pH < 5.8, N(c) attained positive values, approaching 26 for lower ionic strength and pH 3.5. It was also found from the hydrodynamic diameter measurements that for a pH range close to the iep, that is, 4-7, the stability of Fb suspension was very low. These physicochemical characteristics were supplemented by dynamic viscosity measurements, carried out as a function of Fb bulk volume concentration, for various pH values. Using these experimental data the contour length of 80 nm was predicted for Fb molecules in electrolyte solutions. On the other hand, the effective length of the molecule was 53-55 nm for physiological conditions, which suggested a collapsed state of the terminal chains. However, for the range of pH outside the iep, its effective length increased to 65-68 nm. This was interpreted in terms of a significant unfolding of the terminal chains of Fb caused by electrostatic repulsion. The effective charge, contour length, and effective length data derived in this work seem to be the first of this type reported in the literature.
Wasilewska
Monika
M
Institute of Catalysis and Surface Chemistry, Polish Academy of Science, Niezapominajek 8, 30-239 Cracow, Poland. ncwasile@cyf-kr.edu.pl
Adamczyk
Zbigniew
Z
Jachimska
Barbara
B
D000538:Aluminum Silicates; D000818:Animals; D001703:Biophysics / Q000379:methods; D002417:Cattle; D004058:Diffusion; D004563:Electrochemistry / Q000379:methods; D004573:Electrolytes; D004586:Electrophoresis; D005340:Fibrinogen / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007477:Ions; D007700:Kinetics; D008027:Light; D012542:Scattering, Radiation; D013499:Surface Properties
D000538:Aluminum Silicates; D004573:Electrolytes; D007477:Ions; D005340:Fibrinogen; C011934:mica