Coagulant properties of hybrid human/porcine factor VIII molecules.
Human and porcine factor VIII (fVIII) are activated by thrombin to form a heterotrimer composed of subunits designated A1 and A2 derived from the fVIII heavy chain (HC) and a subunit designated A3-C1-C2 derived from the fVIII light chain (LC). Human and porcine fVIII were activated at the same rate to the same peak levels but dissociation of the A2 subunit and concomitant loss of fVIIIa activity at pH 7.4 and 22 degrees C was 3-fold faster with human fVIIIa compared to porcine fVIIIa (0.35 min-1 versus 0.12 min-1, respectively). To determine structural requirements for the increased activity of porcine fVIII, plasma-derived hybrid human/porcine fVIII molecules were isolated. Porcine HC/human LC (pHC/hLC) fVIII had 44-fold higher coagulant activity than reconstituted human fVIII (hHC/hLC), 40-fold higher activity than hHC/pLC, and slightly (1.4-fold) higher activity than reconstituted porcine fVIII (pHC/pLC). Additionally, human and porcine A2 subunits and inactive A1/A3-C1-C2 human and porcine dimers were isolated and reconstitution experiments were done. Addition of the porcine A2 subunit to the human A1/A3-C1-C2 dimer produced coagulant activity similar to that found with porcine fVIIIa and superior to human fVIIIa. These results suggest that human fVIII has weaker coagulant activity than porcine fVIII due to faster dissociation of the A2 subunit and that the A2 subunit itself is responsible for the difference.
Lollar
P
P
Department of Medicine, Emory University, Atlanta, Georgia 30322.
Parker
E T
ET
Fay
P J
PJ
D000818:Animals; D001777:Blood Coagulation; D005169:Factor VIII / Q000378:metabolism*; D015944:Factor VIIIa / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D046911:Macromolecular Substances; D055503:Protein Multimerization; D013552:Swine; D013917:Thrombin / Q000378:metabolism*
D046911:Macromolecular Substances; D015944:Factor VIIIa; D005169:Factor VIII; D013917:Thrombin