Proteomics

Dataset Information

0

Ubiquitin-mediated modulation of necroptotic effector MLKL


ABSTRACT: We aimed to identify lysine (K) residues that are endogenously ubiquitylated upon TNF-induced necroptosis. To this end, we used a quantitative mass spectrometry-based approach. Briefly, MDFs were treated with TSZ or left untreated and protein lysates were digested with trypsin into peptides. Ubiquitylated peptides bearing the di-glycine (K-GG) remnant were enriched by immunoprecipitation with anti-K-ε-GG antibody. A total of three biological replicates were prepared and the 6 samples were labelled with Tandem Mass Tags (TMT). Samples were mixed and analysing by Liquid Chromatography-Tandem Mass Spectroscopy (LC-MS). Quantification was performed to determine the relative abundance of site-specific ubiquitylation events in each condition.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Cell Culture, Fibroblast

SUBMITTER: James Wright  

LAB HEAD: Jyoti Choudhary

PROVIDER: PXD018857 | Pride | 2021-04-13

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
LRG02TMT6plex-V5.msf Msf
LRG02TMT6plex-V5.pdResult Other
LRG02TMT6plex-V5_PeptideGroups.txt Txt
LRG02TMT6plex-V5_Proteins.txt Txt
Lumos50cm_LRG02TMT6plex-P01.raw Raw
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Publications


Necroptosis is a lytic, inflammatory form of cell death that not only contributes to pathogen clearance but can also lead to disease pathogenesis. Necroptosis is triggered by RIPK3-mediated phosphorylation of MLKL, which is thought to initiate MLKL oligomerisation, membrane translocation and membrane rupture, although the precise mechanism is incompletely understood. Here, we show that K63-linked ubiquitin chains are attached to MLKL during necroptosis and that ubiquitylation of MLKL at K219 sig  ...[more]

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