title: Biogenesis of von Willebrand factor-containing organelles in heterologous transfected CV-1 cells.
abstract: Von Willebrand factor (vWF) is a multimeric protein involved in the adhesion of platelets to an injured vessel wall. vWF is synthesized by the endothelial cell and the megakaryocyte as a precursor protein (pro-vWF) that consists of four repeated domains, denoted D1-D2-D'-D3-A1-A2-A3-D4-B1-B2-B3-C1-C2. Previously, we have defined the domains on the pro-vWF molecule involved in dimerization as well as the domains involved in multimer assembly of vWF dimers. In the endothelial cell, part of the vWF multimers is stored in specialized organelles, the Weibel-Palade bodies. By using immunoelectron microscopy, we demonstrate that upon expression of full-length vWF cDNA, vWF-containing organelles are encountered in monkey kidney CV-1 cells that are morphologically similar to the endothelial-specific Weibel-Palade bodies. Expression in CV-1 cells of a series of vWF cDNA deletion mutants, lacking one or more domains, revealed that only those vWF mutant proteins that are able to assemble into multimers are encountered in dense-cored vesicles. Our data show that this process is independent of a particular domain on vWF and indicate that a 'condensed', multimeric vWF is required for targeting to the Weibel-Palade body.
authors.affiliation: Department of Molecular Biology and Blood Coagulation, Central Laboratory, The Netherlands Red Cross Blood Transfusion Service, Amsterdam.
authors.forename: J
authors.initials: J
authors.lastname: Voorberg
authors.forename: R
authors.initials: R
authors.lastname: Fontijn
authors.forename: J
authors.initials: J
authors.lastname: Calafat
authors.forename: H
authors.initials: H
authors.lastname: Janssen
authors.forename: J A
authors.initials: JA
authors.lastname: van Mourik
authors.forename: H
authors.initials: H
authors.lastname: Pannekoek
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D004247:DNA; D005455:Fluorescent Antibody Technique; D000882:Haplorhini; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D009154:Mutation; D015388:Organelles / Q000378:metabolism* / Q000648:ultrastructure; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D014841:von Willebrand Factor / Q000235:genetics / Q000378:metabolism*
chemical_list: D011498:Protein Precursors; D014841:von Willebrand Factor; C050818:prepro-von Willebrand factor; D004247:DNA