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Purification and characterization of human coagulation factor V
The Journal of Biological Chemistry
|January 25, 1981
Summary
Human coagulation Factor V was purified and characterized. Thrombin activation significantly increases its coagulation activity through specific molecular cleavages, revealing intermediate and final products.
Area of Science:
- Biochemistry
- Hematology
- Molecular Biology
Background:
- Human coagulation Factor V (FV) is a critical protein in the blood clotting cascade.
- Understanding FV's structure and activation is essential for hemostasis research.
Purpose of the Study:
- To purify and characterize human coagulation Factor V.
- To investigate the molecular mechanisms of Factor V activation by thrombin and Russell's viper venom.
Main Methods:
- Purification of human FV using a multi-step process including precipitation, adsorption, and chromatography.
- Analysis of FV structure and activation intermediates using SDS-PAGE.
- Assay of coagulation activity following enzymatic treatment.
Main Results:
- Human FV purified to homogeneity is a single polypeptide chain (Mr = 335,000) and a glycoprotein with high sialic acid content.
- Thrombin activation yields a 25-30 fold increase in coagulation activity, involving cleavage of three bonds and generating specific intermediates and products.
- Russell's viper venom activator splits FV into two components, fully activating it and correlating activity with a 95,000-91,000 component.
Conclusions:
- Human FV activation by thrombin involves distinct cleavage patterns and intermediates compared to bovine FV.
- The generation of a ~95 kDa fragment is strongly associated with the functional activation of human FV.