Proteomics

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Global analysis of impact of Atg5 mutation on steady-state protein levels

ABSTRACT: This experiment analyzes the global impact of Atg5-/- mutation on steady-state protein levels by a standard SILAC experiment. WT+vector cells were grown in isotopically labeled media for multiple passages in order to fully label the proteome. Unlabeled Atg5-/- cells and labeled WT+vector cells were grown to confluency and protein extracts from quiescent cultures were combined at a 1:1 ratio and analyzed by LC-MS/MS. In this experiment, ratios of labeled to unlabeled spectra report on changes in steady-state expression levels of proteins induced by impairment of autophagy. Table of files Cells Fractionation Raw Data Filenames Wildtype & Atg5-/- Yes SILAC_(1-8).raw (8 fractions)

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Fibroblast

SUBMITTER: Tian Zhang  

LAB HEAD: Sina Ghaemmaghami

PROVIDER: PXD003558 | Pride | 2016-06-14

REPOSITORIES: Pride

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Publications

Global Analysis of Cellular Protein Flux Quantifies the Selectivity of Basal Autophagy.

Zhang Tian T   Shen Shichen S   Qu Jun J   Ghaemmaghami Sina S  

Cell reports 20160303 10

In eukaryotic cells, macroautophagy is a catabolic pathway implicated in the degradation of long-lived proteins and damaged organelles. Although it has been demonstrated that macroautophagy can selectively degrade specific targets, its contribution to the basal turnover of cellular proteins has not been quantified on proteome-wide scales. In this study, we created autophagy-deficient primary human fibroblasts and quantified the resulting changes in basal degradative flux by dynamic proteomics. O  ...[more]

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