title: The platelet actin cytoskeleton associates with SNAREs and participates in alpha-granule secretion.
abstract: 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 alpha-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 alpha-granule secretion. Inhibition of actin polymerization prevented alpha-granule secretion in this system, and purified platelet actin could substitute for platelet cytosol to support alpha-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 alpha-granule release.
authors.affiliation: Division of Hemostasis and Thrombosis, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215, USA.
authors.forename: Kamil
authors.initials: K
authors.lastname: Woronowicz
authors.forename: James R
authors.initials: JR
authors.lastname: Dilks
authors.forename: Nataliya
authors.initials: N
authors.lastname: Rozenvayn
authors.forename: Louisa
authors.initials: L
authors.lastname: Dowal
authors.forename: Price S
authors.initials: PS
authors.lastname: Blair
authors.forename: Christian G
authors.initials: CG
authors.lastname: Peters
authors.forename: Lucyna
authors.initials: L
authors.lastname: Woronowicz
authors.forename: Robert
authors.initials: R
authors.lastname: Flaumenhaft
mesh_terms: D000199:Actins / Q000276:immunology / Q000378:metabolism; D001792:Blood Platelets / Q000378:metabolism* / Q000502:physiology; D019086:Bridged Bicyclo Compounds, Heterocyclic; D003594:Cytoplasmic Granules / Q000276:immunology / Q000378:metabolism*; D003599:Cytoskeleton / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D017830:Octoxynol / Q000378:metabolism; D015539:Platelet Activation / Q000187:drug effects / Q000276:immunology / Q000502:physiology*; D050765:Qa-SNARE Proteins / Q000276:immunology / Q000378:metabolism; D050600:SNARE Proteins / Q000378:metabolism*; D050827:Syntaxin 1 / Q000378:metabolism; D053778:Thiazolidines; D001620:beta-Thromboglobulin
chemical_list: D000199:Actins; D019086:Bridged Bicyclo Compounds, Heterocyclic; C061615:PPBP protein, human; D050765:Qa-SNARE Proteins; D050600:SNARE Proteins; D050827:Syntaxin 1; D053778:Thiazolidines; D001620:beta-Thromboglobulin; D017830:Octoxynol; C037067:latrunculin A