Characterization of recombinant human factor VIII.

摘要: Recently, complete human factor VIII DNA clones have been obtained and subsequently expressed in baby hamster kidney cells (Wood, W. I., Capon, D. J., Simonsen, C. C., Eaton, L., Gitschier, Keyt, B., Seeburg, P. H., Smith, Hollingshead, P., Wion, K. Delwart, E., Tuddenham, E. G. D., Vehar, A., Lawn, R. M. (1984) Nature 312, 330-337). The recombinant (rVIII) protein secreted from these has now purified allowing its structural analysis comparison to plasma-derived (pdVIII). Analysis of rVIII by sodium dodecyl sulfate-polyacrylamide gel electrophoresis shows that it consists multiple polypeptides with relative mobilities (Mr) ranging 80,000-210,000. same pattern is also observed for pdVIII resolved electrophoresis. proteins associated are recognized antibodies a Western blot. When subjected isoelectric focusing they into similar bands. Thrombin, Xa, activated C, which modulate activity proteolysis, process the manner do pdVIII. As case pdVIII, thrombin activation coagulant correlates generation subunits Mr 73,000, 50,000 43,000. These appear form metal-(perhaps Ca2+) linked complex. EDTA inactivates thrombin-activated being regenerated after addition molar excess MnCl2. results suggest structurally functionally very