Related Experiment Videos
Recombinant human protein C: comparative functional studies with human plasma protein C
R M Madden1, C Oppenheimer, R Wydro
1Lab Service, VA Medical Center, Denver CO 80220.
Thrombosis Research
|February 1, 1990
Summary
Recombinant protein C (r-PC) shows functional similarity to plasma protein C (n-PC). However, r-PC exhibits significantly faster thrombin-thrombomodulin activation, suggesting potential therapeutic advantages in antithrombotic regulation.
Area of Science:
- Biochemistry
- Hematology
- Protein Engineering
Background:
- Protein C (PC) is crucial for antithrombotic regulation; deficiencies are linked to thrombosis.
- Recombinant techniques enable commercial-scale production of human PC for potential therapeutic use.
Purpose of the Study:
- To comparatively investigate the functional properties of recombinant protein C (r-PC) and native plasma protein C (n-PC).
- To assess the potential of r-PC as a therapeutic agent by comparing its activation and activity to n-PC.
Main Methods:
- Immunopurification of both r-PC and n-PC.
- Comparative analysis of Protac C and thrombin-thrombomodulin (T-TM) activation kinetics.
- Evaluation of anticoagulant and profibrinolytic activities of activated PC forms.
Main Results:
- Protac C activation rates and kinetics were identical for r-PC and n-PC.
- Thrombin-thrombomodulin (T-TM) activation of r-PC was significantly more efficient (Kcat/Km = 378) than n-PC (Kcat/Km = 35).
- Activated r-PC and n-PC demonstrated equivalent anticoagulant and profibrinolytic activities.
Conclusions:
- Recombinant protein C (r-PC) possesses comparable functional properties to native plasma protein C (n-PC).
- The enhanced T-TM activation of r-PC may stem from a higher proportion of single-chain PC, which could adopt a more readily activatable conformation.