Investigation of the aggregation and activation of prothrombin using quasi-elastic light scattering
| Autor: | Agarwal Gp, Aune Kc, Gallagher Jg, Armeniades Cd |
|---|---|
| Rok vydání: | 1977 |
| Předmět: |
Aqueous solution
Chemical Phenomena Light Molecular mass Chemistry Physical Macromolecular Substances Chemistry Dimer Analytical chemistry Biochemistry Light scattering Enzyme Activation Molecular Weight chemistry.chemical_compound Thrombin Fragmentation (mass spectrometry) medicine Humans Scattering Radiation Prothrombin Quasi-elastic Light Scattering Mass fraction medicine.drug |
| Zdroj: | Biochemistry. 16:1865-1870 |
| ISSN: | 1520-4995 0006-2960 |
| DOI: | 10.1021/bi00628a016 |
| Popis: | The technique of quasi-elastic light scattering was used to measure the translational diffusion coefficient, D, of purified human prothrombin in buffered aqueous solutions and to monitor for the first time the fragmentation of this protein as it is converted to thrombin. The values of D20,w, measured at two different concentrations, are 4.72 X 10(-7) CM2/S at 2MG/CM3 and 4.51 X 10(-7) CM2/S at 5MG/CM3; the corresponding molecular weights (Mw of 92 000 and 120 000), obtained by combining sedimentation velocity measurements with the diffusion data, confirm the presence of molecular aggregates of prothrombin in these solutions. These results, as well as analysis of the intensity-intensity autocorrelation functions from two-component systems with various dimer conformations, indicated the presence of end-to-end dimers in these prothrombin solutions. The values obtained for D indicate a dimer weight fraction of 0.4 to 0.5 in the 2 mg/cm3 solution and 0.6 or greater in the 5 mg/cm3 solution. The fragmentation of prothrombin was monitored in a nonphysiologic activation system, containing taipan snake venom, dihexanoylphosphatidylcholine, and CaCl2. At a temperature of 15 degrees C, conversion to thrombin proceeded very slowly and was still incomplete after 90 h. A method for determing the percentage of converted prothrombin is an activated system containing aggregates from the average value of D and light scattering data is discussed. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|