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KMID : 0366220150500020103
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Korean Journal of Hematology
2015 Volume.50 No. 2 p.103 ~ p.108
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Human coagulation factor VIII domain-specific recombinant polypeptide expression
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Choi Su-Jin
Jang Ki-Jung
Lim Jeong-A
Kim Hye-Sun
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Abstract
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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.
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KEYWORD
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Hemophilia A, Coagulation factor VIII, Coagulation factor IX, Domain-specific recombinant FVIII, Hep3B hepatocytes
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