title: Human coagulation factor VIII domain-specific recombinant polypeptide expression.
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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Biological Science, College of Natural Sciences, Ajou University, Suwon, Korea.
authors.forename: Su Jin
authors.initials: SJ
authors.lastname: Choi
authors.affiliation: Department of Biological Science, College of Natural Sciences, Ajou University, Suwon, Korea.
authors.forename: Ki Jung
authors.initials: KJ
authors.lastname: Jang
authors.affiliation: National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda, MD, USA.
authors.forename: Jeong-A
authors.initials: JA
authors.lastname: Lim
authors.affiliation: Department of Biological Science, College of Natural Sciences, Ajou University, Suwon, Korea.
authors.forename: Hye Sun
authors.initials: HS
authors.lastname: Kim
keywords: Coagulation factor IX; Coagulation factor VIII; Domain-specific recombinant FVIII; Hemophilia A; Hep3B hepatocytes