title: Platelet secretion and hemostasis require syntaxin-binding protein STXBP5.
abstract: Genome-wide association studies (GWAS) have linked genes encoding several soluble NSF attachment protein receptor (SNARE) regulators to cardiovascular disease risk factors. Because these regulatory proteins may directly affect platelet secretion, we used SNARE-containing complexes to affinity purify potential regulators from human platelet extracts. Syntaxin-binding protein 5 (STXBP5; also known as tomosyn-1) was identified by mass spectrometry, and its expression in isolated platelets was confirmed by RT-PCR analysis. Coimmunoprecipitation studies showed that STXBP5 interacts with core secretion machinery complexes, such as syntaxin-11/SNAP23 heterodimers, and fractionation studies suggested that STXBP5 also interacts with the platelet cytoskeleton. Platelets from Stxbp5 KO mice had normal expression of other key secretory components; however, stimulation-dependent secretion from each of the 3 granule types was markedly defective. Secretion defects in STXBP5-deficient platelets were confirmed via lumi-aggregometry and FACS analysis for P-selectin and LAMP-1 exposure. Interestingly, STXBP5-deficient platelets had altered granule cargo levels, despite having normal morphology and granule numbers. Consistent with secretion and cargo deficiencies, Stxbp5 KO mice showed dramatic bleeding in the tail transection model and defective hemostasis in the FeCl3-induced carotid injury model. Transplantation experiments indicated that these defects were due to loss of STXBP5 in BM-derived cells. Our data demonstrate that STXBP5 is required for normal arterial hemostasis, due to its contributions to platelet granule cargo packaging and secretion.
authors.forename: Shaojing
authors.initials: S
authors.lastname: Ye
authors.forename: Yunjie
authors.initials: Y
authors.lastname: Huang
authors.forename: Smita
authors.initials: S
authors.lastname: Joshi
authors.forename: Jinchao
authors.initials: J
authors.lastname: Zhang
authors.forename: Fanmuyi
authors.initials: F
authors.lastname: Yang
authors.forename: Guoying
authors.initials: G
authors.lastname: Zhang
authors.forename: Susan S
authors.initials: SS
authors.lastname: Smyth
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Li
authors.forename: Yoshimi
authors.initials: Y
authors.lastname: Takai
authors.forename: Sidney W
authors.initials: SW
authors.lastname: Whiteheart
mesh_terms: D000818:Animals; D001792:Blood Platelets / Q000166:cytology* / Q000378:metabolism; D002469:Cell Separation; D005434:Flow Cytometry; D006487:Hemostasis; D006801:Humans; D013058:Mass Spectrometry; D051379:Mice; D018345:Mice, Knockout; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D019007:P-Selectin / Q000378:metabolism; D055503:Protein Multimerization; D050683:R-SNARE Proteins / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D050600:SNARE Proteins / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism
chemical_list: D009419:Nerve Tissue Proteins; D019007:P-Selectin; D050683:R-SNARE Proteins; D050600:SNARE Proteins; C484767:STXBP5 protein, human; C112618:tomosyn protein, mouse