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Thromb Haemost 1999; 82(03): 1177-1181
DOI: 10.1055/s-0037-1614349
Letters to the Editor
Schattauer GmbH

Small GTP-binding Protein RalA Associates with Weibel-Palade Bodies in Endothelial Cells

Hubert P. J. C. de Leeuw
1   From the Department of Blood Coagulation, CLB, Amsterdam, The Netherlands
,
Pauline M. Wijers-Koster
1   From the Department of Blood Coagulation, CLB, Amsterdam, The Netherlands
,
Jan A. van Mourik
1   From the Department of Blood Coagulation, CLB, Amsterdam, The Netherlands
,
Jan Voorberg
1   From the Department of Blood Coagulation, CLB, Amsterdam, The Netherlands
› Author Affiliations
Further Information

Publication History

Received 31 December 1998

Accepted after revision 30 April 1999

Publication Date:
09 December 2017 (online)

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Summary

In endothelial cells von Willebrand factor (vWF) and P-selectin are stored in dense granules, so-called Weibel-Palade bodies. Upon stimulation of endothelial cells with a variety of agents including thrombin, these organelles fuse with the plasma membrane and release their content. Small GTP-binding proteins have been shown to control release from intracellular storage pools in a number of cells. In this study we have investigated whether small GTP-binding proteins are associated with Weibel-Palade bodies. We isolated Weibel-Palade bodies by centrifugation on two consecutive density gradients of Percoll. The dense fraction in which these subcellular organelles were highly enriched, was analysed by SDS-PAGE followed by GTP overlay. A distinct band with an apparent molecular weight of 28,000 was observed. Two-dimensional gel electrophoresis followed by GTP overlay revealed the presence of a single small GTP-binding protein with an isoelectric point of 7.1. A monoclonal antibody directed against RalA showed reactivity with the small GTP-binding protein present in subcellular fractions that contain Weibel-Palade bodies. The small GTPase RalA was previously identified on dense granules of platelets and on synaptic vesicles in nerve terminals. Our observations suggest that RalA serves a role in regulated exocytosis of Weibel-Palade bodies in endothelial cells.

Keywords

Weibel-Palade bodies - RalA - GTP - von Willebrand factor - endothelial cells
 

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