Proteomics

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Acetylation Dynamics and Stoichiometry in Saccharomyces cerevisiae


ABSTRACT: Quantitative MS analysis of acetylation in yeast using SILAC labeling and MaxQuant. Download Index of Raw files first. We used quantitative mass spectrometry to analyze acetylation dynamics and stoichiometry in Saccharomyces cerevisiae. We found that acetylation accumulated in growth-arrested cells in a manner that depended on acetyl-CoA generation in distinct subcellular compartments. We used stable isotope labeling with amino acids in cell culture to quantify differences in protein, acetylation, and phosphorylation abundance by MS. Proteins from whole cell lysates were digested to peptides and acetylated peptides enriched using a polyclonal anti-acetyllysine antibody. Peptide fractions were analyzed by reversed-phase liquid chromatography coupled to high resolution liquid chromatography‐tandem mass spectrometry (LC-MS/MS) and raw MS data were computationally processed using MaxQuant.

INSTRUMENT(S): LTQ Orbitrap Velos, Q Exactive

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

SUBMITTER: Brian Weinert  

PROVIDER: PXD000507 | Pride | 2014-02-03

REPOSITORIES: Pride

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Publications

Acetylation dynamics and stoichiometry in Saccharomyces cerevisiae.

Weinert Brian T BT   Iesmantavicius Vytautas V   Moustafa Tarek T   Schölz Christian C   Wagner Sebastian A SA   Magnes Christoph C   Zechner Rudolf R   Choudhary Chunaram C  

Molecular systems biology 20140130


Lysine acetylation is a frequently occurring posttranslational modification; however, little is known about the origin and regulation of most sites. Here we used quantitative mass spectrometry to analyze acetylation dynamics and stoichiometry in Saccharomyces cerevisiae. We found that acetylation accumulated in growth-arrested cells in a manner that depended on acetyl-CoA generation in distinct subcellular compartments. Mitochondrial acetylation levels correlated with acetyl-CoA concentration in  ...[more]

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