Human coagulation factor VIII domain-specific recombinant polypeptide expression
| Autor: | Hye Sun Kim, Ki Jung Jang, Su Jin Choi, Jeong A Lim |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
congenital
hereditary and neonatal diseases and abnormalities Coagulation factor IX business.industry Domain-specific recombinant FVIII Hep3B hepatocytes Hematology Coagulation Factor IX Bioinformatics Hemophilia A law.invention Coagulation Biochemistry law Hemostasis hemic and lymphatic diseases Coagulation factor VIII Recombinant DNA Human coagulation factor VIII Medicine Original Article business |
| Zdroj: | Blood research |
| ISSN: | 2287-979X |
| Popis: | Background Hemophilia A is caused by heterogeneous mutations in F8. Coagulation factor VIII (FVIII), the product of F8, is composed of multiple domains designated A1-A2-B-A3-C1-C2. FVIII is known to interact with diverse proteins, and this characteristic may be important for hemostasis. However, little is known about domain-specific functions or their specific binding partners. Methods To determine F8 domain-specific functions during blood coagulation, the FVIII domains A1, A2, A3, and C were cloned from Hep3B hepatocytes. Domain-specific recombinant polypeptides were glutathione S-transferase (GST)- or polyhistidine (His)-tagged, over-expressed in bacteria, and purified by specific affinity chromatography. Results Recombinant polypeptides of predicted sizes were obtained. The GST-tagged A2 polypeptide interacted with coagulation factor IX, which is known to bind the A2 domain of activated FVIII. Conclusion Recombinant, domain-specific polypeptides are useful tools to study the domain-specific functions of FVIII during the coagulation process, and they may be used for production of domain-specific antibodies. |
| Databáze: | OpenAIRE |
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