Importance of protease inhibition in studies on purified factor VIII (antihaemophilic factor).
According to some authors factor VIII procoagulant activity may be dissociable from carrier protein (MW approximately 2 X 10(6) by agarose gel filtration, e.g. at high ionic strength. We were able to reproduce this phenomenon. However, addition of protease inhibitor (Trasylol) prevented the appearance of low molecular weight peak of factor VIII procoagulant activity both at high ionic strength and elevated temperature (37 degrees C). We conclude from our results that procoagulant activity and carrier protein (von Willebrand factor, factor VIII antigen) are closely associated functional sites of native factor VIII macromolecule. Consequently, proteolytic degradation should be avoided in functional and structural studies on factor VIII and especially in preparing factor VIII concentrate for therapeutic use.
D000941:Antigens; D007611:Aprotinin; D002352:Carrier Proteins / Q000302:isolation & purification; D005169:Factor VIII / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000502:physiology*; D006801:Humans; D006868:Hydrolysis; D009994:Osmolar Concentration; D011487:Protein Conformation; D013696:Temperature; D014841:von Willebrand Factor
D000941:Antigens; D002352:Carrier Proteins; D014841:von Willebrand Factor; D005169:Factor VIII; D007611:Aprotinin