Multimeric composition of endothelial cell-derived von Willebrand factor.
The multimeric composition of human endothelial cell (EC)-derived von Willebrand factor (vWF) was studied using SDS-agarose gel electrophoresis and autoradiography. Two multimers were found in lysates prepared from confluent cultures of human umbilical vein endothelial cells. The smaller multimer had a molecular weight (mol wt) of approximately 950 Kd, while the second was larger than those seen in plasma. When electrophoresis was performed using the discontinuous buffer system of Ruggeri and Zimmerman, the small multimer consisted of a single band migrating with the slowest-moving component of the corresponding plasma triplet. The large EC-vWF multimer was detected in culture media conditioned with EC monolayers for ten minutes. It remained the only multimer in media conditioned for up to three days. Calcium ionophore A23187 increased the amount of the large vWF multimer released into the culture media, but did not change its multimeric composition. The small multimer was never detected in the EC-conditioned media. These findings suggest that (1) a large, fully polymerized multimer of vWF is released from the ECs, while the small multimer probably represents a major intermediate component in the process of multimerization, and (2) plasma vWF multimers are probably generated from the large endothelial vWF after it is released into the circulation.
Tsai
H M
HM
Division of Hematology, Montefiore Medical Center, Bronx, NY 10467.
Nagel
R L
RL
Hatcher
V B
VB
Sussman
I I
II
D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D046911:Macromolecular Substances; D008970:Molecular Weight; D014471:Umbilical Veins; D014841:von Willebrand Factor / Q000096:biosynthesis / Q000302:isolation & purification*
D046911:Macromolecular Substances; D014841:von Willebrand Factor