PXD011294-2

PXD011294 is an original dataset announced via ProteomeXchange.

Title	Defective AP-3-dependent VAMP8 trafficking impairs Weibel-Palade body exocytosis in Hermansky-Pudlak Syndrome type 2 blood outgrowth endothelial cells.
Description	Weibel-Palade bodies (WPBs) are endothelial secretory organelles that contain VWF, P-selectin and CD63. Release of VWF from WPBs is crucial for platelet adhesion during primary hemostasis. Endosomal trafficking of proteins like CD63 to WPBs during maturation is dependent on the AP-3 complex. Mutations in the AP3B1 gene, which encodes the AP-3 β1 subunit, result in Hermansky-Pudlak syndrome 2 (HPS-2), a rare genetic disorder that leads to neutropenia and a mild bleeding diathesis. This is caused by abnormal granule formation in neutrophils and platelets due to defects in trafficking of cargo to secretory organelles. The impact of these defects on the secretory pathway of the endothelium is largely unknown. In this study we have investigated the role of AP-3-dependent mechanisms in trafficking of proteins during WPB maturation in endothelial cells. An ex vivo patient-derived endothelial model of HPS-2 was established using blood outgrowth endothelial cells (BOECs) that were isolated from an HPS-2 patient with compound heterozygous mutations in AP3B1. HPS-2 BOECs and CRISPR/Cas9-engineered AP3B1-/- BOECs contain WPBs that are entirely devoid of CD63, indicative of disrupted endosomal trafficking to the WPB membrane. HPS-2 BOECs have impaired Ca2+- and cAMP-mediated WPB exocytosis. Whole proteome analysis of HPS-2 BOECs revealed that apart from AP-3β1 also the AP-3µ1 subunit and the v-SNARE VAMP8 were depleted. Stimulus-induced VWF secretion was impaired in CRISPR/Cas9-engineered VAMP8-/- BOECs. Our data show that defects in AP-3 dependent maturation of WPBs impairs WPB exocytosis by affecting the recruitment of the WPB-localized member of the SNARE fusion machinery VAMP8.
HostingRepository	PRIDE
AnnounceDate	2019-01-31
AnnouncementXML	Submission_2019-01-31_06:04:28.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Maartje van den Biggelaar
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue; acetylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion ETD

Revision	Datetime	Status	ChangeLog Entry
0	2018-10-05 08:10:33	ID requested
1	2019-01-31 05:45:18	announced
⏵ 2	2019-01-31 06:04:30	announced	Updated publication reference for PubMed record(s): 30630984.
3	2024-10-22 04:43:44	announced	2024-10-22: Updated project metadata.

Karampini E, Schillemans M, Hofman M, van Alphen F, de Boer M, Kuijpers TW, van den Biggelaar M, Voorberg J, Bierings R, Defective AP-3-dependent VAMP8 trafficking impairs Weibel-Palade body exocytosis in Hermansky-Pudlak Syndrome type 2 blood outgrowth endothelial cells. Haematologica, 104(10):2091-2099(2019) [pubmed]

curator keyword: Biological

submitter keyword: endothelial cells, weibel-palade bodies, AP3 complex, HPS2

M. van den Biggelaar
contact affiliation	Department of Molecular and Cellular Hemostasis, Sanquin Research, Amsterdam, The Netherlands
contact email	m.vandenbiggelaar@sanquin.nl
lab head
Maartje van den Biggelaar
contact affiliation	Sanquin Research
contact email	m.vandenbiggelaar@sanquin.nl
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/01/PXD011294

PRIDE project URI

• PRIDE
  1. PXD011294
     1. Label: PRIDE project
     2. Name: Defective AP-3-dependent VAMP8 trafficking impairs Weibel-Palade body exocytosis in Hermansky-Pudlak Syndrome type 2 blood outgrowth endothelial cells.