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von Willebrand factor biosynthesis and partitioning between constitutive
and regulated pathways of secretion after thrombin stimulation
T Mayadas, DD Wagner and PJ Simpson
Department of Medicine, University of Rochester School of Medicine &
Dentistry, NY 14642.
The major part of von Willebrand factor (vWf) synthesized in cultured
endothelial cells is secreted constitutively without stimulation and
consists of all multimeric forms of vWf. In contrast, stimulation with
secretagogues such as thrombin results in the release of vWf from the
storage pool, the Weibel-Palade bodies which contain only the largest, most
biologically potent multimeric forms of vWf. We wished to determine whether
the signal for release of vWf might also function as a signal for
replenishment of the vWf by enhancing de novo biosynthesis and if
replenishment of the vWf storage pool involved a diversion of newly
synthesized vWf from the constitutive pathway to the regulated pathway. vWf
mRNA and protein levels in unstimulated human umbilical vein endothelial
cells were compared with cells that were briefly stimulated with 1 U/mL
thrombin for 15 minutes and then incubated without thrombin for periods up
to 72 hours. A comparison was also made between unstimulated cells and
cells continuously exposed to thrombin for up to 48 hours. Thrombin
stimulation, brief or continuous, had no significant effect on subsequent
biosynthesis of vWf protein or vWf- specific mRNA. Since thrombin releases
vWf only from the storage pool, we examined the possibility of diversion of
newly synthesized vWf from the constitutive pathway to the regulated
pathway. Cells were pulse- labeled, incubated for 15 minutes with and
without thrombin, chased for various periods in unlabeled media, and
briefly restimulated with thrombin. No significant redistribution of vWf
between the two pathways was observed as a result of thrombin stimulation
for the time periods tested.(ABSTRACT TRUNCATED AT 250 WORDS)
Volume 73,
Issue 3,
pp. 706-711,
02/15/1989
Copyright © 1989 by The American Society of Hematology
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