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Vol. 13, Issue 5, 1582-1593, May 2002

Selective and Signal-dependent Recruitment of Membrane Proteins to Secretory Granules Formed by Heterologously Expressed von Willebrand Factor

Anastasia D. Blagoveshchenskaya,dagger Matthew J. Hannah,* Simon Allen,Dagger and Daniel F. Cutler§

 *Medical Research Council Laboratory for Molecular Cell Biology, Cell Biology Unit, and Department of Biochemistry and Molecular Biology, University College London, London WC1E 6BT, United Kingdom;  dagger Vollum Institute L-474, Oregon Health Sciences University, Portland, Oregon; and  Dagger Academic Unit of Neurology, Division of Genomic Medicine, Medical School, University of Sheffield, Sheffield, S10 2RX, United Kingdom

von Willebrand factor (vWF) is a large, multimeric protein secreted by endothelial cells and involved in hemostasis. When expressed in AtT-20 cells, vWF leads to the de novo formation of cigar-shaped organelles similar in appearance to the Weibel-Palade bodies of endothelial cells in which vWF is normally stored before regulated secretion. The membranes of this vWF-induced organelle, termed the pseudogranule, are uncharacterized. We have examined the ability of these pseudogranules, which we show are secretagogue responsive, to recruit membrane proteins. Coexpression experiments show that the Weibel-Palade body proteins P-selectin and CD63, as well as the secretory organelle membrane proteins vesicle-associated membrane protein-2 and synaptotagmin I are diverted away from the endogenous adrenocorticotropic hormone-containing secretory granules to the vWF-containing pseudogranules. However, transferrin receptor, lysosomal-associated membrane protein 1, and sialyl transferase are not recruited. The recruitment of P-selectin is dependent on a tyrosine-based motif within its cytoplasmic domain. Our data show that vWF pseudogranules specifically recruit a subset of membrane proteins, and that in a process explicitly driven by the pseudogranule content (i.e., vWF), the active recruitment of at least one component of the pseudogranule membrane (i.e., P-selectin) is dependent on residues of P-selectin that are cytosolic and therefore unable to directly interact with vWF.

§ Corresponding author. E-mail address: d.cutler{at}ucl.ac.uk.

Molecular Biology of the Cell
Vol. 13, 1582-1593, May 2002
Copyright © 2002 by The American Society for Cell Biology


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