PXD022979

PXD022979 is an original dataset announced via ProteomeXchange.

Title	Proteomic identification of the UDP-GlcNAc : PI α1-6 GlcNAc-transferase subunits of the glycosylphosphatidylinositol biosynthetic pathway of Trypanosoma brucei.
Description	The first step of glycosylphosphatidylinositol (GPI) anchor biosynthesis in all eukaryotes is the addition of N-acetylglucosamine (GlcNAc) to phosphatidylinositol (PI) which is catalysed by a UDP-GlcNAc : PI α1-6 GlcNAc-transferase. This enzyme has been shown to be a complex of at least seven subunits in mammalian cells and a similar complex of homologous subunits has been postulated in yeast. Homologs of most of these mammalian and yeast subunits were identified in the Trypanosoma brucei predicted protein database. The putative catalytic subunit of the T. brucei complex,TbGPI3, was epitope tagged with three consecutive c-Myc sequences at its C-terminus. Immunoprecipitation of TbGPI3-3Myc followed by native polyacrylamide gel electrophoresis and anti-Myc Western blot showed that it is present in a ~240 kDa complex. Label-free quantitative proteomics were performed to compare anti-Myc pull-downs from lysates of TbGPI-3Myc expressing and wild type cell lines. TbGPI3-3Myc was the most highly enriched protein in the TbGPI3-3Myc lysate pull-down and partner proteins TbGPI15, TbGPI9, TbGPI2, TbGPI1 and TbERI1 were also identified with significant enrichment. Our proteomics data also suggest that an Arv-1 like protein (TbArv1) is a subunit of the T. brucei complex. Yeast and mammalian Arv1 have been previously implicated in GPI biosynthesis, but here we present the first experimental evidence for physical association of Arv1 with GPI biosynthetic machinery. A putative E2-ligase has also been tentatively identified as part of the T. brucei UDP-GlcNAc : PI α1-6 GlcNAc-transferase complex.
HostingRepository	PRIDE
AnnounceDate	2021-02-26
AnnouncementXML	Submission_2021-02-26_08:40:01.339.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Michele Tinti
SpeciesList	scientific name: Trypanosoma brucei; NCBI TaxID: 5691;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive Plus

Revision	Datetime	Status	ChangeLog Entry
0	2020-12-07 08:55:35	ID requested
⏵ 1	2021-02-26 08:40:01	announced

Dataset with its publication pending

submitter keyword: Trypanosoma brucei, glycosylphosphatidylinositol, GlcNAc-transferase, TbArv1

Mike Ferguson
contact affiliation	Biological Chemistry and Drug Discovery Wellcome Centre for Anti-Infectives Research (WCAIR)
contact email	m.a.j.ferguson@dundee.ac.uk
lab head
Michele Tinti
contact affiliation	Dundee University
contact email	m.tinti@dundee.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD022979
     1. Label: PRIDE project
     2. Name: Proteomic identification of the UDP-GlcNAc : PI α1-6 GlcNAc-transferase subunits of the glycosylphosphatidylinositol biosynthetic pathway of Trypanosoma brucei.