2 FUNCTIONALLY DISTINCT POOLS OF VITRONECTIN (VN) IN THE BLOOD-CIRCULATION - IDENTIFICATION OF A HEPARIN-BINDING COMPETENT POPULATION OF VNWITHIN PLATELET ALPHA-GRANULES

The biological functions of vitronectin (Vn) are dependent on its conf ormation. Whereas plasma Vn is present in a conformation that does not bind to heparin, platelet Vn has been recognized to be in a multimeri c, conformationally altered form. To further understand the characteri stics of platelet Vn, the molecules present in plasma and total and si ze-fractionated platelet releasates were compared (1) immunologically using three conformationally sensitive epitope-defined monoclonal anti bodies, (2) functionally for their ability to interact with heparin, a nd (3) structurally using denaturing and nondenaturing polyacrylamide gel electrophoresis (PAGE). Our data indicate that Vn is present in pl atelet releasates in two molecular weight (M(r)) forms. The high M(r) fractions contain conformationally and structurally altered Vn capable of interacting with heparin, and this form is distinct from plasma Vn and purified denatured Vn. In contrast, the lower M(r) forms of Vn ar e similar to plasma Vn. To determine if the presence of multimeric Vn requires platelet activation, platelets were disintegrated by sonicati on and fractionated by density gradients. Combined sodium dodecyl sulf ate-PAGE (SDS-PAGE) and immunoblotting analysis showed a codistributio n of multimeric Vn and type 1 plasminogen activator inhibitor in alpha -granule-rich fractions. Thus, platelet Vn is stored in a structurally and functionally distinct form from the molecule in plasma, raising t he possibility that platelet-derived heparin-binding competent Vn will accumulate in areas of vascular injury. (C) 1996 by The American Soci ety of Hematology.