PXD037158

PXD037158 is an original dataset announced via ProteomeXchange.

Title	O-fucosylation of Thrombospondin Type I Repeats on Thrombospondin 1 is dispensable for trafficking to Platelets
Description	Thrombospondin 1 (THBS1) is a secreted extracellular matrix glycoprotein that regulates a variety of cellular and physiological processes. THBS1’s diverse functions are attributed to interactions between the modular domains of THBS1 with an array of proteins found in the extracellular matrix. THBS1’s three Thrombospondin Type 1 Repeats (TSRs) domains are modified with O-linked glucose-fucose disaccharide and C-mannose. It is unknown whether these modifications impact trafficking and/or function of THBS1 in vivo. The O-fucose is added by Protein O-fucosyltransferase 2 (POFUT2) and is sequentially extended to the disaccharide by β3glucosyltransferase (B3GLCT). The C-mannose is added by one or more of four C-mannosyltransferases. O-fucosylation by POFUT2/B3GLCT in the endoplasmic reticulum has been proposed to play a role in quality control by locking TSR domains into their 3-dimensional fold, allowing for proper secretion of many O-fucosylated substrates. Prior studies showed the siRNA knockdown of POFUT2 in HEK293T cells blocked secretion of TSRs 1-3 from THBS1. Here we demonstrated that secretion of THBS1 TSRs 1-3 was not reduced by CRISPR-Cas9-mediated knockout of POFUT2 in HEK293T cells and demonstrated that knockout of Pofut2 or B3glct in mice did not reduce trafficking of endogenous THBS1 to platelet secretory granules, a major site of THBS1 expression. Additionally, we demonstrated that all three TSRs from platelet THBS1 were highly C-mannosylated, which has been shown to stabilize TSRs in vitro. Combined these results suggested that POFUT2 substrates with TSRs that are also modified by C-mannose may be less susceptible to trafficking defects resulting from loss of the glucose-fucose disaccharide.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_07:31:34.472.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Steven Berardinelli
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	monohydroxylated residue; deamidated residue; iodoacetamide derivatized residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2022-10-03 11:26:09	ID requested
1	2023-02-02 12:16:46	announced
⏵ 2	2023-11-14 07:31:39	announced	2023-11-14: Updated project metadata.

Berardinelli SJ, Sillato AR, Grady RC, Neupane S, Ito A, Haltiwanger RS, Holdener BC, O-fucosylation of thrombospondin type I repeats is dispensable for trafficking thrombospondin 1 to platelet secretory granules. Glycobiology, 33(4):301-310(2023) [pubmed]

submitter keyword: POFUT2/O-fucosylation/C-mannosylation/Thrombospondin 1/protein trafficking

Robert S. Haltiwanger
contact affiliation	Associate Professor, Biochemistry and Molecular Biology Department, University of Georgia, Athens, GA, United States of American
contact email	rhalti@uga.edu
lab head
Steven Berardinelli
contact affiliation	University of Georgia
contact email	sjb96455@uga.edu
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/2023/02/PXD037158

PRIDE project URI

• PRIDE
  1. PXD037158
     1. Label: PRIDE project
     2. Name: O-fucosylation of Thrombospondin Type I Repeats on Thrombospondin 1 is dispensable for trafficking to Platelets