Proteomics

Dataset Information

0

Proteomic identification of the UDP-GlcNAc : PI α1-6 GlcNAc-transferase subunits of the glycosylphosphatidylinositol biosynthetic pathway of Trypanosoma brucei.


ABSTRACT: 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.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Trypanosoma Brucei

SUBMITTER: Michele Tinti  

LAB HEAD: Mike Ferguson

PROVIDER: PXD022979 | Pride | 2021-02-26

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
ZJ-sm1-1.raw Raw
ZJ-sm1-2.raw Raw
ZJ-sm1-3.raw Raw
ZJ-sm1-4.raw Raw
ZJ-sm2-1.raw Raw
Items per page:
1 - 5 of 10

Similar Datasets

2021-01-28 | PXD023124 | Pride
2013-08-23 | E-GEOD-48284 | biostudies-arrayexpress
2013-08-23 | GSE48284 | GEO
2018-09-24 | PXD006635 | Pride
2010-08-01 | E-GEOD-22337 | biostudies-arrayexpress
2019-05-21 | GSE121472 | GEO
2019-05-21 | GSE112667 | GEO
2017-07-18 | GSE84502 | GEO
2018-10-22 | PXD009763 | Pride
2017-07-20 | GSE101642 | GEO