STRUCTURAL-ANALYSIS OF RECOMBINANT VON-WILLEBRAND-FACTOR - IDENTIFICATION OF HETERO-DIMERS AND HOMO-DIMERS

Wild-type full-length cDNA of von Willebrand factor (VWF) was expresse d in CHO cells. Recombinant VWF (rvWF) was obtained and its molecular composition investigated by two-dimensional electrophoresis. Results o f first dimension electrophoresis under non-reducing conditions showed that rvWF-dimer represents a mixture of three different species. Seco nd dimension electrophoresis under reducing conditions revealed, that these protein species represent (i) homo-dimers of either two unproces sed or two fully processed, mature vWF polypeptides, and (ii) the hete ro-dimer of unprocessed and mature rvWF monomers. Compared with vWF-di mers from human plasma, which contained predominantly proteolytically degraded polypeptides, recombinant vWF-dimers were shown to consist of non-proteolyzed subunits only.