Biosynthesis of von Willebrand protein by human megakaryocytes.
Immunofluorescence staining of buffy coat smears from a patient with chronic myelogenous leukemia in accelerated phase showed that approximately 13% of all nucleated cells contained von Willebrand protein and, therefore, appeared to be of megakaryocytic origin. This was confirmed by positive staining with antisera against platelet factor 4 and platelet glycoproteins. Short-term cultures of the buffy coat, which lacked endothelial cells, were metabolically labeled with [35S]methionine, and von Willebrand protein was immunopurified from cell lysates and culture medium. Cultures from this patient synthesized and secreted von Willebrand protein, in contrast with cultures from other patients with leukemia, who lacked circulating megakaryocytes, and from normal volunteers. The subunit composition of the megakaryocytic von Willebrand protein was very similar to that of human umbilical vein endothelial cells. The size of the processed subunit (220 kD) and of the cellular (260 kD) and secreted (275 kD) precursors from the two cell types were indistinguishable by gel electrophoresis. Furthermore, the ratio of precursor to processed subunit and the pattern of cellular and secreted nonreduced multimers were very similar. It appears, therefore, that the processing steps in biosynthesis of von Willebrand protein used by the megakaryocytes are very similar to those of umbilical vein endothelial cells.
L A
LA
Sporn
S I
SI
Chavin
V J
VJ
Marder
D D
DD
Wagner
D000328:Adult; D002478:Cells, Cultured; D004727:Endothelium / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D007951:Leukemia, Myeloid / Q000097:blood*; D008297:Male; D008533:Megakaryocytes / Q000378:metabolism*; D008970:Molecular Weight; D013194:Staining and Labeling; D014471:Umbilical Veins; D014841:von Willebrand Factor / Q000096:biosynthesis* / Q000302:isolation & purification
D014841:von Willebrand Factor