Cloning and Recombinant Expression of Mouse Coagulation Factor X

Engineering of recombinant coagulation factor X variants, which can be activated by tumor-associated proteinases may lead to the development of new therapeutic molecules. However, the evaluation of such variants requires an appropriate animal model. Therefore, we isolated the complete coding sequenc...

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Bibliographic Details
Published in:Thrombosis research 1998-10, Vol.92 (1), p.33-41
Main Authors: Heidtmann, Hans-Heinrich, Kontermann, Roland E.
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Base Sequence
Biological and medical sciences
Blood coagulation
Blood Coagulation - drug effects
Blood coagulation. Blood cells
Cattle
cDNA
Cell Line
Cloning, Molecular
Coagulation factors
Conserved Sequence
DNA, Complementary - genetics
Factor X
Factor X - genetics
Factor X - isolation & purification
Factor X - pharmacology
Fundamental and applied biological sciences. Psychology
Gene Expression
Genetic Variation
Humans
In Vitro Techniques
Mice
Molecular and cellular biology
Molecular Sequence Data
Rabbits
Recombinant expression
Recombinant Proteins - genetics
Recombinant Proteins - isolation & purification
Recombinant Proteins - pharmacology
Sequence Homology, Amino Acid
Serine proteinase
Species Specificity
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Summary:Engineering of recombinant coagulation factor X variants, which can be activated by tumor-associated proteinases may lead to the development of new therapeutic molecules. However, the evaluation of such variants requires an appropriate animal model. Therefore, we isolated the complete coding sequence of mouse coagulation factor X from mouse liver cDNA by polymerase chain reaction. The deduced amino acid sequence codes for a prepro protein of 481 amino acids homologous to factor X sequences from various species. Recombinant mouse factor X was expressed in human embryonic kidney cells and secreted into cell culture supernatant as zymogen, which could be converted to catalytically active factor Xa by Russell’s viper venom. Purified recombinant mouse factor X restored coagulation in human factor X deficient plasma, demonstrating that mouse factor X is able to functionally interact with the human blood coagulation system. Recombinant mouse factor X opens the possibility to analyze therapeutically useful variants in the mouse system.
ISSN:0049-3848
1879-2472
DOI:10.1016/S0049-3848(98)00110-8