Human coagulation factor VIII domain-specific recombinant polypeptide expression.
- Author:
Su Jin CHOI
1
;
Ki Jung JANG
;
Jeong A LIM
;
Hye Sun KIM
Author Information
- Publication Type:Original Article
- Keywords: Hemophilia A; Coagulation factor VIII; Coagulation factor IX; Domain-specific recombinant FVIII; Hep3B hepatocytes
- MeSH: Antibodies; Bacteria; Blood Coagulation; Chromatography, Affinity; Clone Cells; Factor IX; Factor VIII*; Glutathione Transferase; Hemophilia A; Hemostasis; Hepatocytes; Humans; Peptides
- From:Blood Research 2015;50(2):103-108
- CountryRepublic of Korea
- Language:English
- Abstract: 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.
