Structural analysis of recombinant von Willebrand factor: identification of hetero- and homo-dimers.
Wild-type full-length cDNA of von Willebrand factor (vWF) was expressed in CHO cells. Recombinant vWF (rvWF) was obtained and its molecular composition investigated by two-dimensional electrophoresis. Results of first dimension electrophoresis under non-reducing conditions showed that rvWF-dimer represents a mixture of three different species. Second dimension electrophoresis under reducing conditions revealed, that these protein species represent (i) homo-dimers of either two unprocessed or two fully processed, mature vWF polypeptides, and (ii) the hetero-dimer of unprocessed and mature rvWF monomers. Compared with vWF-dimers from human plasma, which contained predominantly proteolytically degraded polypeptides, recombinant vWF-dimers were shown to consist of non-proteolyzed subunits only.
Fischer
B
B
IMMUNO AG, Biomedical Research Center, Donau, Austria.
Mitterer
A
A
Schlokat
U
U
DenBouwmeester
R
R
Dorner
F
F
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry; D014841:von Willebrand Factor / Q000737:chemistry*
D011994:Recombinant Proteins; D014841:von Willebrand Factor