Polymerization and gelation of fibrinogen in D2O.
The solution properties of fibrinogen and the thrombin-induced activation and gelation of fibrinogen in 95% D2O at pH 7.4 were compared to those in H2O under similar conditions. The initial release rates of fibrinopeptides A and B in D2O were slightly slower than those in H2O. However, the values of the Michaelis-Menten parameters Km and V for the release of the two peptides in D2O and H2O in the presence of 0.5 M NaCl were about the same. From turbidity measurements at 450 nm it is obvious that fibrinogen is soluble in a slightly more narrow range of NaCl concentration and that the fibrin gels have a higher degree of lateral aggregation in D2O than in H2O. The variation of fibrinogen concentration, thrombin concentration, pH and ionic a strength have a similar dependence on the final gel structure and clotting time in D2O and H2O. SDS-gel electrophoresis on fibrin samples, which were cross-linked by factor XIII, yielded results where the cross-linking of the gamma-chain appeared to be the same in D2O and H2O. The alpha-chain cross-linking was somewhat faster in D2O than in H2O. When fibrinogen solutions in 95% D2O were incubated at 20 mM CaCl2, a slow gelation of fibrinogen was observed, which was found to be induced by trace amounts of factor XIII. The final gel turbidity appeared to be about the same for this gelation as for that induced by thrombin. The differences in solubility for fibrinogen, kinetics for the enzyme reaction and optical properties for the fibrin gels in D2O and H2O may be explained by differences in electrostatic interactions, hydrogen bonding and hydration of fibrinogen in these two media.
Larsson
U
U
Department of Coagulation Research, Karolinska Institute, Stockholm, Sweden.
D002122:Calcium Chloride; D003903:Deuterium; D004563:Electrochemistry; D004591:Electrophoresis, Polyacrylamide Gel; D005340:Fibrinogen; D005344:Fibrinopeptide A; D005345:Fibrinopeptide B; D005782:Gels; D006801:Humans; D006860:Hydrogen Bonding; D006863:Hydrogen-Ion Concentration; D046911:Macromolecular Substances; D009391:Nephelometry and Turbidimetry; D011108:Polymers; D012965:Sodium Chloride; D014867:Water
D005782:Gels; D046911:Macromolecular Substances; D011108:Polymers; D014867:Water; D005344:Fibrinopeptide A; D005345:Fibrinopeptide B; D012965:Sodium Chloride; D005340:Fibrinogen; D003903:Deuterium; D002122:Calcium Chloride