Blood Res.  2015 Jun;50(2):103-108. 10.5045/br.2015.50.2.103.

Human coagulation factor VIII domain-specific recombinant polypeptide expression

• Choi SJ
• Jang KJ
• Lim JA
• Kim HS

• KMID: 2172774
• DOI: http://doi.org/10.5045/br.2015.50.2.103

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.

Keyword

Hemophilia A; Coagulation factor VIII; Coagulation factor IX; Domain-specific recombinant FVIII; Hep3B hepatocytes

MeSH Terms

Antibodies
Bacteria
Blood Coagulation
Chromatography, Affinity
Clone Cells
Factor IX
Factor VIII*
Glutathione Transferase
Hemophilia A
Hemostasis
Hepatocytes
Humans
Peptides
Antibodies
Factor IX
Factor VIII
Glutathione Transferase
Peptides