title: Functional self-association of von Willebrand factor during platelet adhesion under flow.
abstract: We have used recombinant wild-type human von Willebrand factor (VWF) and deletion mutants lacking the A1 and A3 domains, as well as specific function-blocking monoclonal antibodies, to demonstrate a functionally relevant self-association at the interface of soluble and surface-bound VWF. Platelets perfused at the wall shear rate of 1,500 s(-1) over immobilized VWF lacking A1 domain function failed to become tethered to the surface when they were in a plasma-free suspension with erythrocytes, but adhered promptly if soluble VWF with functional A1 domain was added to the cells. The same results were observed when VWF was immobilized onto collagen through its A3 domain and soluble VWF with deleted A3 domain was added to the cells. Thus, VWF bound to glass or collagen sustains a process of homotypic self-association with soluble VWF multimers that, as a result, can mediate platelet adhesion. The latter finding demonstrates that direct immobilization on a substrate is not a strict requirement for VWF binding to platelet glycoprotein Ibalpha. The dynamic and reversible interaction of surface-bound and soluble VWF appears to be specifically homotypic, because immobilized BSA, human fibrinogen, and fibronectin cannot substitute for VWF in the process. Our findings highlight a newly recognized role of circulating VWF in the initiation of platelet adhesion. The self-assembly of VWF multimers on an injured vascular surface may provide a relevant contribution to the arrest of flowing platelets opposing hemodynamic forces, thus facilitating subsequent thrombus growth.
authors.affiliation: Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, CA 92037, USA.
authors.forename: Brian
authors.initials: B
authors.lastname: Savage
authors.forename: Jan J
authors.initials: JJ
authors.lastname: Sixma
authors.forename: Zaverio M
authors.initials: ZM
authors.lastname: Ruggeri
mesh_terms: D001792:Blood Platelets / Q000166:cytology; D002448:Cell Adhesion; D003094:Collagen / Q000378:metabolism; D004492:Edetic Acid / Q000494:pharmacology; D004912:Erythrocytes / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D017353:Gene Deletion; D006439:Hemodynamics; D006801:Humans; D018715:Microscopy, Video; D010974:Platelet Aggregation; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism; D014841:von Willebrand Factor / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D014841:von Willebrand Factor; D003094:Collagen; D004492:Edetic Acid