Dataset Information

Syntaxin-3 is a Weibel-Palade body localized SNARE protein that regulates Von Willebrand factor secretion from endothelial cells

ABSTRACT: Endothelial cells store von Willebrand factor (VWF) in rod-shaped secretory organelles, called Weibel-Palade bodies (WPBs). WPB exocytosis is coordinated by a complex network of Rab GTPases, Rab-effectors and SNARE proteins. We have previously identified STXBP1 as the link between the Rab27A-Slp4-a complex on WPBs and the SNARE proteins syntaxin-2 and -3. In this study we investigate the function of syntaxin-3 in VWF secretion. In human umbilical vein endothelial cells (HUVECs) and in blood outgrowth endothelial cells (BOECs) from healthy controls endogenous syntaxin-3 immunolocalized to WPBs. A detailed analysis of BOECs isolated from a patient with variant microvillus inclusion disease (MVID), carrying a homozygous mutation in STX3 (STX3-/-), showed a loss of syntaxin-3 protein and absence of WPB-associated syntaxin-3 immunoreactivity. Ultrastructural analysis revealed no detectable differences in morphology or prevalence of immature or mature WPBs in control versus STX3-/- BOECs. VWF multimer analysis showed normal patterns in plasma of the MVID patient, and media from STX3-/- BOECs, together indicating WPB formation and maturation are unaffected by absence of syntaxin-3. However, a clear defect in Ca2+ and cAMP-mediated VWF secretion was found in the STX3-/- BOECs. Co-immunoprecipitation studies showed that syntaxin-3 associates with the WPB SNAREs SNAP23 and VAMP8. Our data reveal syntaxin-3 as a novel WPB-associated SNARE controlling VWF secretion and highlight the complex regulation of WPB exocytosis by multiple SNARE complexes.

INSTRUMENT(S): Orbitrap Fusion ETD

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Endothelial Cell, Blood

SUBMITTER: Bart van den Eshof

LAB HEAD: Maartje van den Biggelaar

PROVIDER: PXD006176 | Pride | 2018-07-04

REPOSITORIES: Pride

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Publications

Weibel-Palade Body Localized Syntaxin-3 Modulates Von Willebrand Factor Secretion From Endothelial Cells.

Schillemans Maaike M Karampini Ellie E van den Eshof Bart L BL Gangaev Anastasia A Hofman Menno M van Breevoort Dorothee D Meems Henriët H Janssen Hans H Mulder Aat A AA Jost Carolina R CR Escher Johanna C JC Adam Rüdiger R Carter Tom T Koster Abraham J AJ van den Biggelaar Maartje M Voorberg Jan J Bierings Ruben R

Arteriosclerosis, thrombosis, and vascular biology 20180607 7

<h4>Objective</h4>Endothelial cells store VWF (von Willebrand factor) in rod-shaped secretory organelles, called Weibel-Palade bodies (WPBs). WPB exocytosis is coordinated by a complex network of Rab GTPases, Rab effectors, and SNARE (soluble NSF attachment protein receptor) proteins. We have previously identified STXBP1 as the link between the Rab27A-Slp4-a complex on WPBs and the SNARE proteins syntaxin-2 and -3. In this study, we investigate the function of syntaxin-3 in VWF secretion.<h4>App ...[more]

PMID: 29880488

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