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Protein C activation by recombinant thrombomodulin in plasma
T Edano1, N Komine, H Yoshizaki
1Tokyo Research Laboratories, Kowa Co., Ltd., Japan.
Biological & Pharmaceutical Bulletin
|March 26, 1998
Summary
Glycosaminoglycan-modified urinary thrombomodulin (GAG-UTM) enhances protein C activation more effectively than unmodified UTM. This modified protein demonstrates potent anticoagulant properties in plasma, offering therapeutic potential.
Area of Science:
- Biochemistry
- Molecular Biology
- Thrombosis Research
Background:
- Human urinary thrombomodulin (UTM) plays a role in regulating coagulation.
- Recombinant UTM (r-UTM) has been developed, but its efficacy can be limited.
- Glycosaminoglycan (GAG) modification is a strategy to enhance protein function.
Purpose of the Study:
- To investigate the properties of recombinant GAG-modified UTM (GAG-UTM).
- To compare the protein C activation and thrombin inhibition activities of GAG-UTM and r-UTM.
- To determine if GAG-UTM accelerates protein C activation in plasma.
Main Methods:
- Expression of GAG-UTM in C127 cells.
- In vitro assays measuring protein C activation by thrombin and thrombin inhibition by antithrombin III (ATIII) in buffer.
- Western blot analysis using anti-protein C antibody to detect activated protein C generation in plasma.
Main Results:
- GAG-UTM exhibited more potent acceleration of protein C activation and thrombin inhibition by ATIII than r-UTM in buffer.
- GAG-UTM suppressed thrombin generation in plasma more effectively than r-UTM during protein C activation.
- Western blot confirmed greater generation of activated protein C by GAG-UTM compared to r-UTM in plasma.
Conclusions:
- GAG-UTM demonstrates enhanced anticoagulant activity compared to unmodified r-UTM.
- The acceleration of activated protein C formation by GAG-UTM is effective in a plasma environment.
- GAG-UTM shows promise as a therapeutic agent for thrombotic disorders.