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Identification of and partial characterization of platelet vitronectin:
evidence for complex formation with platelet-derived plasminogen activator
inhibitor-1
KT Preissner, S Holzhuter, C Justus and G Muller-Berghaus
Clinical Research Unit for Blood Coagulation and Thrombosis of the Max-
Planck-Gesellschaft Justus-Liebig-Universitat, Giessen, FRG.
Vitronectin (VN; = complement S-protein), a plasma glycoprotein that is
also associated with extracellular sites, was identified in washed human
platelets contaminated with less than 0.05% of plasma VN. A specific
enzyme-linked immunosorbent assay (ELISA) for VN has been developed and was
used to detect and to quantitate VN in detergent extracts of washed
platelets with 8.1 +/- 4.6 micrograms/10(9) platelets (n = 10),
representing about 0.8% of the plasma VN pool. Platelet and plasma VN were
similar by immunochemical criteria using Western-blot analysis, although
platelet VN was mainly found as partially proteolyzed polypeptide. Total
release of platelet VN occurred at optimal doses of Ca-ionophore 23187 or
thrombin, whereas no VN was released by platelet treatment with digitonin
or Staphylococcus alpha-toxin. During stimulation of washed platelets with
various concentrations of thrombin, the nearly concomitant release of VN
and plasminogen activator inhibitor-1 (PAI-1) together with platelet factor
4 indicated the association of VN with inner-platelet storage granules.
Furthermore, platelet VN and PAI-1 in Ca-ionophore releasates comigrated
during ultracentrifugation in high mol wt fractions of sucrose density
gradients, indicating a possible association of both components. Complex
formation of platelet VN and PAI-1 was verified by a sensitive
enzyme-linked immunosorbent assay (ELISA) and accounts at least in part for
a high molecular form of platelet VN. The identification of platelet VN and
its binding to platelet PAI-1 raises the possibility that VN, in contrast
to other adhesive proteins, may participate in localized regulatory
functions of blood coagulation and fibrinolysis in platelet-matrix
interactions and the protection of the matrix against proteolysis.
Volume 74,
Issue 6,
pp. 1989-1996,
11/01/1989
Copyright © 1989 by The American Society of Hematology
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