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Small GTP-binding protein RalA associates with Weibel-Palade bodies in endothelial cells.
de Leeuw, H P; Wijers-Koster, P M; van Mourik, J A; Voorberg, J.
Afiliación
  • de Leeuw HP; Department of Blood Coagulation, CLB, Amsterdam, The Netherlands.
Thromb Haemost ; 82(3): 1177-81, 1999 Sep.
Article en En | MEDLINE | ID: mdl-10494784
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.
Asunto(s)
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Endotelio Vascular / Cuerpos de Weibel-Palade / Proteínas de Unión al GTP ral / GTP Fosfohidrolasas Tipo de estudio: Prognostic_studies / Risk_factors_studies Límite: Humans Idioma: En Revista: Thromb Haemost Año: 1999 Tipo del documento: Article País de afiliación: Países Bajos Pais de publicación: Alemania
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Endotelio Vascular / Cuerpos de Weibel-Palade / Proteínas de Unión al GTP ral / GTP Fosfohidrolasas Tipo de estudio: Prognostic_studies / Risk_factors_studies Límite: Humans Idioma: En Revista: Thromb Haemost Año: 1999 Tipo del documento: Article País de afiliación: Países Bajos Pais de publicación: Alemania