The Platelet Actin Cytoskeleton Associates with SNAREs and Participates in α-Granule Secretion

• Published in: Biochemistry (Easton) 2010-06, Vol.49 (21), p.4533-4542
• Main Authors: Woronowicz, Kamil, Dilks, James R, Rozenvayn, Nataliya, Dowal, Louisa, Blair, Price S, Peters, Christian G, Woronowicz, Lucyna, Flaumenhaft, Robert
• Format: Article
• Language: English
• Subjects: Actins - immunology; Actins - metabolism; beta-Thromboglobulin; Blood Platelets - metabolism; Blood Platelets - physiology; Blood Platelets - secretion; Bridged Bicyclo Compounds, Heterocyclic; Cytoplasmic Granules - immunology; Cytoplasmic Granules - metabolism; Cytoplasmic Granules - secretion; Cytoskeleton - immunology; Cytoskeleton - metabolism; Cytoskeleton - physiology; Humans; Octoxynol - metabolism; Platelet Activation - drug effects; Platelet Activation - immunology; Platelet Activation - physiology; Qa-SNARE Proteins - immunology; Qa-SNARE Proteins - metabolism; SNARE Proteins - metabolism; Syntaxin 1 - metabolism; Thiazolidines
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi100541t

Following platelet activation, platelets undergo a dramatic shape change mediated by the actin cytoskeleton and accompanied by secretion of granule contents. While the actin cytoskeleton is thought to influence platelet granule secretion, the mechanism for this putative regulation is not known. We found that disruption of the actin cytoskeleton by latrunculin A inhibited α-granule secretion induced by several different platelet agonists without significantly affecting activation-induced platelet aggregation. In a cell-free secretory system, platelet cytosol was required for α-granule secretion. Inhibition of actin polymerization prevented α-granule secretion in this system, and purified platelet actin could substitute for platelet cytosol to support α-granule secretion. To determine whether SNAREs physically associate with the actin cytoskeleton, we isolated the Triton X-100 insoluble actin cytoskeleton from platelets. VAMP-8 and syntaxin-2 associated only with actin cytoskeletons of activated platelets. Syntaxin-4 and SNAP-23 associated with cytoskeletons isolated from either resting or activated platelets. When syntaxin-4 and SNAP-23 were tested for actin binding in a purified protein system, only syntaxin-4 associated directly with polymerized platelet actin. These data show that the platelet cytoskeleton interacts with select SNAREs and that actin polymerization facilitates α-granule release.