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[Procedures for the isolation and characterization of human antithrombin III]
Behring Institute Mitteilungen
|February 1, 1986
Summary
Antithrombin III, a key inhibitor of blood clotting factors, was purified and characterized. Heparin acts as a catalyst by forming a complex with Antithrombin III and activated coagulation factors.
Area of Science:
- Biochemistry
- Hematology
- Protein chemistry
Context:
- Antithrombin III (AT III) is the primary inhibitor of activated coagulation factors.
- Understanding AT III's function is crucial for hemostasis research.
- Human plasma is the source for AT III isolation.
Purpose:
- To isolate and biochemically characterize human Antithrombin III.
- To elucidate the mechanism of inhibition of coagulation factors by AT III.
- To explore the role of heparin in AT III-mediated anticoagulation.
Summary:
- Human Antithrombin III was purified using affinity chromatography and characterized for uniformity via HPLC and PAA gradient gel electrophoresis.
- Biochemical properties determined include a molecular weight of 70,000 daltons and an isoelectric point of pH 4.5.
- Kinetic studies confirmed competitive inhibition of factor Xa by AT III, functioning as a "dead end" inhibitor, with heparin potentially acting as a catalyst via ternary complex formation.
Impact:
- Provides detailed biochemical data on purified human Antithrombin III.
- Clarifies the inhibitory mechanism of AT III and the catalytic role of heparin.
- Discusses the therapeutic implications of Antithrombin III concentrates in coagulation disorders.