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Prepublished online as a Blood First Edition Paper on June 5, 2003; DOI 10.1182/blood-2003-02-0599. This Article Full Text Full Text (PDF) All Versions of this Article: 2003-02-0599v1 102/7/2452    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Michaux, G. Articles by Cutler, D. F. Search for Related Content PubMed PubMed Citation Articles by Michaux, G. Articles by Cutler, D. F. Related Collections Hemostasis, Thrombosis, and Vascular Biology Signal Transduction Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  Blood, 1 October 2003, Vol. 102, No. 7, pp. 2452-2458 HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY Analysis of intracellular storage and regulated secretion of 3 von Willebrand disease–causing variants of von Willebrand factor Grégoire Michaux, Lindsay J. Hewlett, Sarah L. Messenger, Anne C. Goodeve, Ian R. Peake, Martina E. Daly, and Daniel F. Cutler From the Medical Research Council (MRC) Laboratory for Molecular Cell Biology and Cell Biology Unit, University College London, London; and the Academic Unit of Haematology, Division of Genomic Medicine, Royal Hallamshire Hospital, Sheffield, United Kingdom. The rapid exocytosis of von Willebrand factor (VWF) in response to vascular injury can be attributed to the fact that VWF is stored in the Weibel-Palade bodies (WPBs) of endothelial cells. We describe a system for examining the ability of VWF to drive both the formation of a storage compartment and the function of that compartment with respect to regulated secretion. Transient transfection of HEK293 cells with wild-type human VWF cDNA leads to the formation of numerous elongated organelles that resemble WPBs. These "pseudo-WPBs" exhibit the internal structure, as well as the ability to recruit membrane proteins including P-selectin, of bona fide WPBs. Finally, VWF was efficiently secreted upon stimulation by phorbol ester. We used this system to examine 3 VWF mutations leading to von Willebrand disease that affect VWF multimerization and constitutive secretion. Surprisingly we find that all 3 mutants can, to some extent, make pseudo-WPBs that recruit appropriate membrane proteins and that are responsive to secretagogues. The most striking defects are a delay in formation and a reduction in the length and number of pseudo-WPBs in proportion to the clinical severity of the mutation. Studies of pseudo-WPB formation in this system thus yield insights into the structure-function relationships underpinning the ability of VWF to form functional WPBs. (Blood. 2003;102:2452-2458) CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? This article has been cited by other articles: M. van den Biggelaar, A. B. Meijer, J. Voorberg, and K. Mertens Intracellular cotrafficking of factor VIII and von Willebrand factor type 2N variants to storage organelles Blood, March 26, 2009; 113(13): 3102 - 3109. [Abstract] [Full Text] [PDF] D. J. Metcalf, T. D. Nightingale, H. L. Zenner, W. W. Lui-Roberts, and D. F. Cutler Formation and function of Weibel-Palade bodies J. Cell Sci., January 1, 2008; 121(1): 19 - 27. [Abstract] [Full Text] [PDF] G. Michaux, T. J. Pullen, S. L. Haberichter, and D. F. Cutler P-selectin binds to the D'-D3 domains of von Willebrand factor in Weibel-Palade bodies Blood, May 15, 2006; 107(10): 3922 - 3924. [Abstract] [Full Text] [PDF] M. G. Rondaij, R. Bierings, A. Kragt, J. A. van Mourik, and J. Voorberg Dynamics and Plasticity of Weibel-Palade Bodies in Endothelial Cells Arterioscler. Thromb. Vasc. Biol., May 1, 2006; 26(5): 1002 - 1007. [Abstract] [Full Text] [PDF] W. W.Y. Lui-Roberts, L. M. Collinson, L. J. Hewlett, G. Michaux, and D. F. Cutler An AP-1/clathrin coat plays a novel and essential role in forming the Weibel-Palade bodies of endothelial cells J. Cell Biol., August 15, 2005; 170(4): 627 - 636. [Abstract] [Full Text] [PDF] S. L. Haberichter, E. P. Merricks, S. A. Fahs, P. A. Christopherson, T. C. Nichols, and R. R. Montgomery Re-establishment of VWF-dependent Weibel-Palade bodies in VWD endothelial cells Blood, January 1, 2005; 105(1): 145 - 152. [Abstract] [Full Text] [PDF] M. G. Rondaij, E. Sellink, K. A. Gijzen, J. P. ten Klooster, P. L. Hordijk, J. A. van Mourik, and J. Voorberg Small GTP-Binding Protein Ral Is Involved in cAMP-Mediated Release of von Willebrand Factor From Endothelial Cells Arterioscler. Thromb. Vasc. Biol., July 1, 2004; 24(7): 1315 - 1320. [Abstract] [Full Text] [PDF]

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